Pyrrolopyrazine kinase inhibitors

ABSTRACT

The present invention relates to the use of novel pyrrolopyrazine derivatives of Formula I, 
     
       
         
         
             
             
         
       
     
     wherein the variables Q and R are defined as described herein, which inhibit JAK and SYK and are useful for the treatment of auto-immune and inflammatory diseases.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is entitled to the benefit of U.S. provisional patentapplications Ser. No. 61/031,035 filed on Feb. 25, 2008 and Ser. No.61/146,496 filed on Jan. 22, 2009, the disclosures of which areincorporated herein by reference.

FIELD OF THE INVENTION

The present invention relates to the use of novel pyrrolopyrazinederivatives which are JAK and SYK inhibitors and selectively inhibitJAK3 and are useful for the treatment of auto-immune and inflammatorydiseases.

BACKGROUND OF THE INVENTION

Protein kinases constitute one of the largest families of human enzymesand regulate many different signaling processes by adding phosphategroups to proteins; particularly tyrosine kinases phosphorylate proteinson the alcohol moiety of tyrosine residues. The tyrosine kinase familyincludes members that control cell growth, migration, anddifferentiation. Abnormal kinase activity has been implicated in avariety of human diseases including cancers, autoimmune and inflammatorydiseases. Since protein kinases are among the key regulators of cellsignaling they provide a means to modulate cellular function with smallmolecule inhibitors of kinase activity and thus make good drug designtargets. In addition to treatment of kinase-mediated disease processes,selective and efficacious inhibitors of kinase activity are also usefulfor investigation of cell signaling processes and identification ofother cellular targets of therapeutic interest.

The JAKs (JAnus Kinases) are a family of cytoplasmic protein tyrosinekinases including JAK1, JAK2, JAK3 and TYK2. Each of the JAKs ispreferentially associated with the intracytoplasmic portion of discretecytokine receptors (Annu. Rev. Immunol. 16 (1998), pp. 293-322). TheJAKs are activated following ligand binding and initiate signaling byphosphorylating cytokine receptors that, per se, are devoid of intrinsickinase activity. This phosphorylation creates docking sites on thereceptors for other molecules known as STAT proteins (signal transducersand activators of transcription) and the phosphorylated JAKs bindvarious STAT proteins. STAT proteins, or STATs, are DNA binding proteinsactivated by phosphorylation of tyrosine residues, and function both assignaling molecules and transcription factors and ultimately bind tospecific DNA sequences present in the promoters of cytokine-responsivegenes (Leonard et al., (2000), J. Allergy Clin. Immunol. 105:877-888).

JAK/STAT signaling has been implicated in the mediation of many abnormalimmune responses such as allergies, asthma, autoimmune diseases such astransplant (allograft) rejection, rheumatoid arthritis, amyotrophiclateral sclerosis and multiple sclerosis, as well as in solid andhematologic malignancies such as leukemia and lymphomas.

Thus, the JAKs and STATs are components of multiple potentiallyintertwined signal-transduction pathways (Oncogene 19 (2000), pp.5662-5679), which indicates the difficulty of specifically targeting oneelement of the JAK-STAT pathway without interfering with other signaltransduction pathways.

The JAK kinases, including JAK3, are abundantly expressed in primaryleukemic cells from children with acute lymphoblastic leukemia, the mostcommon form of childhood cancer, and studies have correlated STATactivation in certain cells with signals regulating apoptosis (Demoulinet al., (1996), Mol. Cell. Biol. 16:4710-6; Jurlander et al., (1997),Blood. 89:4146-52; Kaneko et al., (1997), Clin. Exp. Immun. 109:185-193;and Nakamura et al., (1996), J. Biol. Chem. 271: 19483-8). They are alsoknown to be important to lymphocyte differentiation, function andsurvival. JAK3 in particular plays an essential role in the function oflymphocytes, macrophages, and mast cells. Given the importance of thisJAK kinase, compounds which modulate the JAK pathway, including thoseselective for JAK3, can be useful for treating diseases or conditionswhere the function of lymphocytes, macrophages, or mast cells isinvolved (Kudlacz et al., (2004) Am. J. Transplant 4:51-57; Changelian(2003) Science 302:875-878). Conditions in which targeting of the JAKpathway or modulation of the JAK kinases, particularly JAK3, arecontemplated to be therapeutically useful include, leukemia, lymphoma,transplant rejection (e.g., pancreas islet transplant rejection, bonemarrow transplant applications (e.g., graft-versus-host disease),autoimmune diseases (e.g., diabetes), and inflammation (e.g., asthma,allergic reactions). Conditions which can benefit for inhibition of JAK3are discussed in greater detail below.

However, in contrast to the relatively ubiquitous expression of JAK1,JAK2 and Tyk2, JAK3 has a more restricted and regulated expression.Whereas some JAKs (JAK1, JAK2, Tyk2) are used by a variety of cytokinereceptors, JAK3 is used only by cytokines that contain a γc in theirreceptor. JAK3, therefore, plays a role in cytokine signaling forcytokines which receptor was shown to date to use the common gammachain; IL-2, IL-4, IL-7, IL-9, IL-15 and IL-21. JAK1 interacts with,among others, the receptors for cytokines IL-2, IL-4, IL-7, IL-9 andIL-21, while JAK2 interacts with, among others, the receptors for IL-9and TNF-alpha. Upon the binding of certain cytokines to their receptors(e.g., IL-2, IL-4, IL-7, IL-9, IL-15 and IL-21), receptoroligomerization occurs, resulting in the cytoplasmic tails of associatedJAK kinases being brought into proximity and facilitating thetrans-phosphorylation of tyrosine residues on the JAK kinase. Thistrans-phosphorylation results in the activation of the JAK kinase.

Animal studies have suggested that JAK3 not only plays a critical rolein B and T lymphocyte maturation, but that JAK3 is constitutivelyrequired to maintain T cell function. Modulation of immune activitythrough this novel mechanism can prove useful in the treatment of T cellproliferative disorders such as transplant rejection and autoimmunediseases.

In particular, JAK3 has been implicated in a variety of biologicalprocesses. For example, the proliferation and survival of murine mastcells induced by IL-4 and IL-9 have been shown to be dependent on JAK3-and gamma chain-signaling (Suzuki et al., (2000), Blood 96:2172-2180).JAK3 also plays a crucial role in IgE receptor-mediated mast celldegranulation responses (Malaviya et al., (1999), Biochem. Biophys. Res.Commun. 257:807-813), and inhibition of JAK3 kinase has been shown toprevent type I hypersensitivity reactions, including anaphylaxis(Malaviya et al., (1999), J. Biol. Chem. 274:27028-27038). JAK3inhibition has also been shown to result in immune suppression forallograft rejection (Kirken, (2001), Transpl. Proc. 33:3268-3270). JAK3kinases have also been implicated in the mechanism involved in early andlate stages of rheumatoid arthritis (Muller-Ladner et al., (2000), J.Immunal. 164:3894-3901); familial amyotrophic lateral sclerosis (Trieuet al., (2000), Biochem Biophys. Res. Commun. 267:22-25); leukemia(Sudbeck et al., (1999), Clin. Cancer Res. 5:1569-1582); mycosisfungoides, a form of T-cell lymphoma (Nielsen et al., (1997), Prac.Natl. Acad. Sci. USA 94:6764-6769); and abnormal cell growth (Yu et al.,(1997), J. Immunol. 159:5206-5210; Catlett-Falcone et al., (1999),Immunity 10:105-115).

JAK3 inhibitors are useful therapy as immunosuppressive agents for organtransplants, xeno transplantation, lupus, multiple sclerosis, rheumatoidarthritis, psoriasis, Type I diabetes and complications from diabetes,cancer, asthma, atopic dermatitis, autoimmune thyroid disorders,ulcerative colitis, Crohn's disease, Alzheimer's disease, Leukemia andother indications where immunosuppression would be desirable.

Non-hematopoietic expression of JAK3 has also been reported, althoughthe functional significance of this has yet to be clarified (J. Immunol.168 (2002), pp. 2475-2482). Because bone marrow transplants for SCID arecurative (Blood 103 (2004), pp. 2009-2018), it seems unlikely that JAK3has essential non-redundant functions in other tissues or organs. Hence,in contrast with other targets of immunosuppressive drugs, therestricted distribution of JAK3 is appealing. Agents that act onmolecular targets with expression limited to the immune system mightlead to an optimal efficacy:toxicity ratio. Targeting JAK3 would,therefore, theoretically offer immune suppression where it is needed(i.e. on cells actively participating in immune responses) withoutresulting in any effects outside of these cell populations. Althoughdefective immune responses have been described in various STAT^(−/−)strains (J. Investig. Med. 44 (1996), pp. 304-311; Curr. Opin. CellBiol. 9 (1997), pp. 233-239), the ubiquitous distribution of STATs andthe fact that those molecules lack enzymatic activity that could betargeted with small-molecule inhibitors has contributed to theirnon-selection as key targets for immunosuppression.

SYK (Spleen Tyrosine Kinase) is a non-receptor tyrosine kinase that isessential for B-cell activation through BCR signaling. SYK becomeactivated upon binding to phosphoryated BCR and thus initiates the earlysignling events following BCR activation. Mice deficient in SYK exhibitan early block in B-cell development (Cheng et al. Nature 378:303, 1995;Turner et al. Nature 378:298, 1995). Therefore inhibition of SYKenzymatic activity in cells is proposed as a treatment for autoimmunedisease through its effects on autoantibody production.

In addition to the role of SYK in BCR signaling and B-cell activation,it also plays a key role in Fc8RI mediated mast cell degranulation andeosinophil activation. Thus, SYK is implicated in allergic disordersincluding asthma (reviewed in Wong et al. Expert Opin Investig Drugs13:743, 2004). SYK binds to the phosphorylated gamma chain of Fc8RI viaits SH2 domains and is essential for downstream signaling (Taylor et al.Mol. Cell. Biol. 15:4149, 1995). SYK deficient mast cells demonstratedefective degranulation, arachidonic acid and cytokine secretion(Costello et al. Oncogene 13:2595, 1996). This also has been shown forpharmacologic agents that inhibit SYK activity in mast cells (Yamamotoet al. J Pharmacol Exp Ther 306:1174, 2003). Treatment with SYKantisense oligonucleotides inhibits antigen-induced infiltration ofeosinophils and neutrophils in an animal model of asthma (Stenton et al.J Immunol 169:1028, 2002). SYK deficient eosinophils also show impairedactivation in response to Fc8R stimulation (Lach-Trifilieffe et al.Blood 96:2506, 2000). Therefore, small molecule inhibitors of SYK willbe useful for treatment of allergy-induced inflammatory diseasesincluding asthma.

In view of the numerous conditions that are contemplated to benefit bytreatment involving modulation of the JAK and/or SYK pathways it isimmediately apparent that new compounds that modulate JAK and/or SYKpathways and methods of using these compounds should provide substantialtherapeutic benefits to a wide variety of patients. Provided herein arenovel pyrrolopyrazine derivatives for use in the treatment of conditionsin which targeting of the JAK and/or SYK pathways or inhibition of JAKor SYK kinases, particularly JAK3, and are therapeutically useful forthe treatment of auto-immune and inflammatory diseases.

SUMMARY OF THE INVENTION

The novel pyrrolopyrazine derivatives provided herein selectivelyinhibit JAK3 and are useful for the treatment of auto-immune andinflammatory diseases. The compounds of the invention modulate the JAKand/or SYK pathways and are useful novel pyrrolopyrazine derivatives forthe treatment of auto-immune and inflammatory diseases, whereinpreferred compounds selectively inhibit JAK3. For example, the compoundsof the invention may inhibit JAK3 and SYK, wherein preferred compoundsare selective for JAK3 of the JAK kinases and are useful novelpyrrolopyrazine derivatives for the treatment of auto-immune andinflammatory diseases. Furthermore, the compounds of the invention mayinhibit JAK3 and JAK2, wherein preferred compounds are selective forJAK3 of the JAK kinases, and are useful novel pyrrolopyrazinederivatives for the treatment of auto-immune and inflammatory diseases.Similarly, the compounds of the invention may inhibit JAK3 and JAK1,wherein preferred compounds are selective for JAK3 of the JAK kinases,and are useful novel pyrrolopyrazine derivatives for the treatment ofauto-immune and inflammatory diseases.

The application provides a compound of Formula I

-   R is R¹, R², R³, or R⁴;    -   R¹ is lower alkyl, lower alkoxy, phenyl, benzyl, heteroaryl,        cycloalkyl, heterocycloalkyl, or cycloalkylalkyl, optionally        substituted with one or more R^(1a);        -   R^(1a) is R^(1b) or R^(1c);            -   R^(1b) is halogen, oxo, hydroxy, or —CN;            -   R^(1c) is —C(═O)O(R^(1f)), —C(═O)CH₂(R^(1e)),                —S(R^(1f)), —S(O)₂(R^(1f)), or —S(═O) (R^(1f)), lower                alkyl, lower alkoxy, amino, amido, lower haloalkyl,                phenyl, heteroaryl, cycloalkyl, heterocycloalkyl,                cycloalkyloxy, or heterocycloalkyloxy optionally                substituted with one or more R^(1d);                -   R^(1d) is H, halogen, hydroxy, lower alkyl, lower                    alkoxy, or lower haloalkyl;                -   R^(1e) is H, lower alkyl, lower alkoxy, —CN, lower                    haloalkyl, phenyl, heteroaryl, cycloalkyl, or                    heterocycloalkyl;                -   R^(1f) is H, lower alkyl, lower haloalkyl, phenyl,                    heteroaryl, cycloalkyl, or heterocycloalkyl;    -   R² is N(R^(2a))₂;        -   each R^(2a) is independently H or R^(2b);            -   each R^(2b) is independently lower alkyl, phenyl,                heteroaryl, cycloalkyl, heterocycloalkyl, or                heterocycloalkyl alkylene, optionally substituted with                one or more R^(2c);                -   R^(2c) is R^(2d) or R^(2e);                -    R^(2d) is halogen, oxo, or hydroxy;                -    R^(2e) is —N(R^(2g))₂, —C(═O)(R^(2g)),                    —C(═O)O(R^(2g)), —C(═O)N(R^(2g))₂,                    —N(R^(2g))C(═O)(R^(2g)), —S(═O)₂(R^(2g)),                    —S(O)₂N(R^(2g))₂, lower alkyl, lower alkoxy, lower                    haloalkyl, phenyl, heteroaryl, heteroaryloxy,                    cycloalkyl, or heterocycloalkyl, optionally                    substituted with one or more R^(2f);                -    each R^(2f) is independently H, halogen, lower                    alkyl, lower alkoxy, lower haloalkyl;                -    each R^(2g) is independently H, lower alkyl, lower                    alkoxy, lower haloalkyl, or phenyl;    -   R³ is —C(═O)R^(3a);        -   R^(3a) is lower alkyl, lower alkoxy, phenyl, or N(R^(3b))₂;            -   each R^(3b) is independently H or lower alkyl;    -   R⁴ is —O(R^(4a));        -   R^(4a) is H or R^(4b);            -   R^(4b) is lower alkyl, phenyl, benzyl, lower haloalkyl,                cycloalkyl, heterocycloalkyl, heteroaryl, optionally                substituted with one or more R^(4c);                -   R^(4c) is halogen, hydroxy, lower alkyl, lower                    haloalkyl, or lower alkoxy;-   Q³ is —O-Q^(3a), —S-Q^(3a), —C(═O)(Q^(3a)),    —O(CH₂)_(m)C(═O)(Q^(3a)), —S(═O)(Q^(3a)), —S(═O)₂(Q^(3a)),    —N(Q^(3a))₂, —N(Q^(3a))S(═O)₂(Q^(3a)), —N(Q^(3a))C(═O)(Q^(3a)),    —C(═O)N(Q^(3a))₂, or —N(Q^(3a))C(═O)N(Q^(3a))₂;    -   each Q^(3a) is independently Q^(3b) or Q^(3c);        -   m is 0, 1, or 2;        -   Q^(3b) is H;        -   Q^(3c) is lower alkyl, lower haloalkyl, phenyl, cycloalkyl,            heterocycloalkyl, or heteroaryl, optionally substituted with            one or more Q^(3d); and            -   each Q^(3d) is independently Q^(3d) is Q^(3e) or Q^(3f);            -   Q^(3e) is halogen or hydroxy;            -   Q^(3f) is lower alkyl, lower alkoxy, lower haloalkyl,                phenyl, cycloalkyl, heterocycloalkyl, or heteroaryl,                optionally substituted with one or more Q^(3g); and                -   each Q^(3g) is independently halogen, hydroxy, lower                    alkyl, lower hydroxyalkyl, lower haloalkyl, or lower                    alkoxy;                    or a pharmaceutically acceptable salt thereof.

In one variation of the above embodiment, R is R¹.

In one variation of the above embodiment, R¹ is lower alkyl.

In one variation of the above embodiment, R¹ is tert-butyl.

In another variation of the above embodiment, R¹ is tert-butyl, Q is Q¹,Q^(1a) is Q^(1c), Q^(1c) is Q^(1e), Q^(1e) is Q^(1e′), and Q^(1e′) ispyrrolidine.

In another variation of the above embodiment, R¹ is —CHC(CH₃)₃.

In another variation of the above embodiment, R¹ is iso-butyl.

In another variation of the above embodiment, R¹ is iso-propyl.

In one embodiment of the compound of Formula I, R¹ is cycloalkyl.

In one embodiment of the compound of Formula I, R¹ is heterocycloalkyl.

In one embodiment of the compound of Formula I, R¹ is benzyl.

In one embodiment of the compound of Formula I, R¹ is phenyl.

In one embodiment of the compound of Formula I, R is R².

In one embodiment of the compound of Formula I, R is R² and R² isNH(R^(2a)).

In one variation of the above embodiment, R^(2a) is R^(2b).

In one variation of the above embodiment, R^(2b) is lower alkyl.

In one variation of the above embodiment, R^(2b) is iso-propyl.

In one embodiment of the compound of Formula I, R^(2b) isheterocycloalkyl.

In one embodiment of the compound of Formula I, R^(2b) is cycloalkyl.

In one embodiment of the compound of Formula I, R^(2b) isheterocycloalkyl alkylene.

In one variation of the above embodiment, R^(2b) is pyrrolidinylmethylene.

In one variation of the above embodiment, R^(2b) is pyrrolidinylmethylene.

The application provides a compound of Formula I selected from the groupconsisting of:

-   2-(Cyclopentyl-methyl-amino)-5H-pyrrolo[2,3-b]pyrazine-7-carboxylic    acid isopropylamide;-   1-[7-(2,2-Dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-2,2-dimethyl-propan-1-one;-   1-(2-Cyclopentyloxy-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one;-   N-[7-(2,2-Dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-benzamide;-   Cyclohexanecarboxylic acid    [7-(2,2-dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-amide;-   N-[7-(2,2-Dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-N-methyl-benzamide;-   N-[7-(2,2-Dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-benzenesulfonamide;-   N-[7-(2,2-Dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-N-methyl-benzenesulfonamide;-   7-(2,2-Dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazine-2-carboxylic    acid dimethylamide;-   7-(2,2-Dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazine-2-carboxylic    acid isopropylamide;-   1-(2-Acetyl-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one;-   1-(2-Isobutyryl-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one;-   2,2-Dimethyl-1-[2-(2-methyl-benzoyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-propan-1-one;-   1-(2-Benzoyl-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one;-   7-(2,2-Dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazine-2-carboxylic    acid cyclopentylamide;-   2,2-Dimethyl-1-[2-(pyridine-4-carbonyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-propan-1-one;-   2,2-Dimethyl-1-[2-(pyridine-3-carbonyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-propan-1-one;-   1-(2-Cycloheptyloxy-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one;-   1-(2-Cyclohexyloxy-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one;-   1-(2-Isopropoxy-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one;-   1-(2-Ethoxy-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one;-   2,2-Dimethyl-1-[2-(2-morpholin-4-yl-2-oxo-ethoxy)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-propan-1-one;-   1-[2-(1,2-Dimethyl-propoxy)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-one;-   1-(2-Isobutoxy-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one;-   1-[2-(1-Benzyl-1H-[1,2,3]triazol-4-yl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-one;-   1-Cyclohexyl-3-[7-(2,2-dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-urea;-   1-Cycloheptyl-3-[7-(2,2-dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-urea;-   2,2-Dimethyl-1-[2-(methyl-phenyl-amino)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-propan-1-one;-   1-(2-sec-Butoxy-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one;-   2,2-Dimethyl-1-(2-phenylamino-5H-pyrrolo[2,3-b]pyrazin-7-yl)-propan-1-one;-   1-(2-Cyclopentyloxy-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one;-   1-[2-(Cyclopentyl-methyl-amino)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-one;-   2,2-Dimethyl-1-(2-phenoxy-5H-pyrrolo[2,3-b]pyrazin-7-yl)-propan-1-one;-   2,2-Dimethyl-1-{2-[4-(4-methyl-piperazin-1-yl)-phenylamino]-5H-pyrrolo[2,3-b]pyrazin-7-yl}-propan-1-one;-   1-Cyclohexyl-3-[7-(2,2-dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-urea;-   1-Cycloheptyl-3-[7-(2,2-dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-urea;-   1-[7-(2,2-Dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-2,2-dimethyl-propan-1-one;    and-   1-[2-(6-Methoxy-pyridin-2-ylamino)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-one.

In one aspect, the application provides a method for treating aninflammatory and/or autoimmune condition comprising administering to apatient in need thereof a therapeutically effective amount of thecompound of Formula I.

In one variation of the above method, the above method further comprisesadministering an additional therapeutic agent selected from achemotherapeutic or anti-proliferative agent, an anti-inflammatoryagent, an immunomodulatory or immunosuppressive agent, a neurotrophicfactor, an agent for treating cardiovascular disease, an agent fortreating diabetes, or an agent for treating immunodeficiency disorders.

In one aspect, the application provides a method for treating aninflammatory condition comprising administering to a patient in needthereof a therapeutically effective amount of the compound of Formula I,wherein R is R².

In one aspect, the application provides a method for inhibiting T-cellproliferative disorder comprising administering to a patient in needthereof a therapeutically effective amount of the compound of Formula I.

In one aspect, the application provides a method for inhibiting T-cellproliferative disorder comprising administering to a patient in needthereof a therapeutically effective amount of the compound of Formula I,wherein R is R².

In one variation of the above method, the proliferative disorder iscancer.

In one aspect, the application provides a method for treating a B-cellproliferative disorder comprising administering to a patient in needthereof a therapeutically effective amount of the compound of Formula I.

In one aspect, the application provides a method for treating an immunedisorder including lupus, multiple sclerosis, rheumatoid arthritis,psoriasis, Type I diabetes, complications from organ transplants, xenotransplantation, diabetes, cancer, asthma, atopic dermatitis, autoimmunethyroid disorders, ulcerative colitis, Crohn's disease, Alzheimer'sdisease, and Leukemia, comprising administering to a patient in needthereof a therapeutically effective amount of the compound of Formula I.

In one aspect, the application provides a method for preventing ortreating all forms of organ rejection, including acute allograft orxenograft rejection and chronic allograft or xenograft rejection, ofvascularized or non-vascularized transplants, comprising administeringto a patient in need thereof a therapeutically effective amount of thecompound of Formula I.

In one aspect, the application provides a method for inhibiting JAK3activity comprising administering the compound of Formula I, wherein thecompound exhibits an IC₅₀ of 50 micromolar or less in an in vitrobiochemical assay of JAK3 activity.

In one variation of the above method, the compound exhibits an IC₅₀ of100 nanomolar or less in an in vitro biochemical assay of JAK3 activity.

In one variation of the above method, the compound exhibits an IC₅₀ of10 nanomolar or less in an in vitro biochemical assay of JAK3 activity.

In one aspect, the application provides a method for inhibiting SYKactivity comprising administering the compound of Formula I, wherein thecompound exhibits an IC₅₀ of 50 micromolar or less in an in vitrobiochemical assay of SYK activity.

In one variation of the above method, the compound exhibits an IC₅₀ of100 nanomolar or less in an in vitro biochemical assay of SYK activity.

In one variation of the above method, the compound exhibits an IC₅₀ of10 nanomolar or less in an in vitro biochemical assay of SYK activity.

In one aspect, the application provides a method for treating aninflammatory condition comprising co-administering to a patient in needthereof an anti-inflammatory compound in combination with atherapeutically effective amount of the compound of Formula I.

In one aspect, the application provides a method for treating an immunedisorder comprising co-administering to a patient in need thereof animmunosuppressant compound in combination with a therapeuticallyeffective amount of the compound of Formula I.

The application provides a pharmaceutical composition comprising thecompound of Formula I, admixed with at least one pharmaceuticallyacceptable carrier, excipient or diluent.

In one variation, the above pharmaceutical composition further comprisesan additional therapeutic agent selected from a chemotherapeutic oranti-proliferative agent, an anti-inflammatory agent, animmunomodulatory or immunosuppressive agent, a neurotrophic factor, anagent for treating cardiovascular disease, an agent for treatingdiabetes, and an agent for treating immunodeficiency disorders.

In one aspect, the application provides a use of the compound of FormulaI in the manufacture of a medicament for the treatment of aninflammatory disorder.

In one aspect, the application provides a use of the compound of FormulaI in the manufacture of a medicament for the treatment of an autoimmunedisorder.

DETAILED DESCRIPTION OF THE INVENTION

The phrase “a” or “an” entity as used herein refers to one or more ofthat entity; for example, a compound refers to one or more compounds orat least one compound. As such, the terms “a” (or “an”), “one or more”,and “at least one” can be used interchangeably herein.

The phrase “as defined herein above” refers to the broadest definitionfor each group as provided in the Summary of the Invention or thebroadest claim. In all other embodiments provided below, substituentswhich can be present in each embodiment and which are not explicitlydefined retain the broadest definition provided in the Summary of theInvention.

As used in this specification, whether in a transitional phrase or inthe body of the claim, the terms “comprise(s)” and “comprising” are tobe interpreted as having an open-ended meaning. That is, the terms areto be interpreted synonymously with the phrases “having at least” or“including at least”. When used in the context of a process, the term“comprising” means that the process includes at least the recited steps,but may include additional steps. When used in the context of a compoundor composition, the term “comprising” means that the compound orcomposition includes at least the recited features or components, butmay also include additional features or components.

As used herein, unless specifically indicated otherwise, the word “or”is used in the “inclusive” sense of “and/or” and not the “exclusive”sense of “either/or”.

The term “independently” is used herein to indicate that a variable isapplied in any one instance without regard to the presence or absence ofa variable having that same or a different definition within the samecompound. Thus, in a compound in which R″ appears twice and is definedas “independently carbon or nitrogen”, both R″s can be carbon, both R″scan be nitrogen, or one R″ can be carbon and the other nitrogen.

When any variable (e.g., R^(2a) or Q^(3g)) occurs more than one time inany moiety or formula depicting and describing compounds employed orclaimed in the present invention, its definition on each occurrence isindependent of its definition at every other occurrence. Also,combinations of substituents and/or variables are permissible only ifsuch compounds result in stable compounds.

The symbols “*” at the end of a bond or

drawn through a bond each refer to the point of attachment of afunctional group or other chemical moiety to the rest of the molecule ofwhich it is a part. Thus, for example:

A bond drawn into ring system (as opposed to connected at a distinctvertex) indicates that the bond may be attached to any of the suitablering atoms.

The term “optional” or “optionally” as used herein means that asubsequently described event or circumstance may, but need not, occur,and that the description includes instances where the event orcircumstance occurs and instances in which it does not. For example,“optionally substituted” means that the optionally substituted moietymay incorporate a hydrogen or a substituent.

The phrase “come together to form a bicyclic ring system” as used hereinmeans join to form a bicyclic ring system, wherein each ring may be madeup of either 4-7 carbon atoms or 4-7 carbon and heteroatoms, and may besaturated or unsaturated.

The term “about” is used herein to mean approximately, in the region of,roughly, or around. When the term “about” is used in conjunction with anumerical range, it modifies that range by extending the boundariesabove and below the numerical values set forth. In general, the term“about” is used herein to modify a numerical value above and below thestated value by a variance of 20%.

The definitions described herein may be appended to formchemically-relevant combinations, such as “heteroalkylaryl,”“haloalkylheteroaryl,” “arylalkylheterocyclyl,” “alkylcarbonyl,”“alkoxyalkyl,” “cycloalkylalkyl” and the like. When the term “alkyl” isused as a suffix following another term, as in “phenylalkyl,” or“hydroxyalkyl,” this is intended to refer to an alkyl group, as definedabove, being substituted with one to two substituents selected from theother specifically-named group. Thus, for example, “phenylalkyl” refersto an alkyl group having one to two phenyl substituents, and thusincludes benzyl, phenylethyl, and biphenyl. An “alkylaminoalkyl” is analkyl group having one to two alkylamino substituents. “Hydroxyalkyl”includes 2-hydroxyethyl, 2-hydroxypropyl,1-(hydroxymethyl)-2-methylpropyl, 2-hydroxybutyl, 2,3-dihydroxybutyl,2-(hydroxymethyl), 3-hydroxypropyl, and so forth. Accordingly, as usedherein, the term “hydroxyalkyl” is used to define a subset ofheteroalkyl groups defined below. The term -(ar)alkyl refers to eitheran unsubstituted alkyl or an aralkyl group. The term (hetero)aryl or(het)aryl refers to either an aryl or a heteroaryl group.

Compounds of formula I may exhibit tautomerism. Tautomeric compounds canexist as two or more interconvertable species. Prototropic tautomersresult from the migration of a covalently bonded hydrogen atom betweentwo atoms. Tautomers generally exist in equilibrium and attempts toisolate an individual tautomers usually produce a mixture whose chemicaland physical properties are consistent with a mixture of compounds. Theposition of the equilibrium is dependent on chemical features within themolecule. For example, in many aliphatic aldehydes and ketones, such asacetaldehyde, the keto form predominates while; in phenols, the enolform predominates. Common prototropic tautomers include keto/enol(—C(═O)—CH-8-C(—OH)═CH—), amide/imidic acid (—C(═O)—NH-8-C(—OH)═N—) andamidine (—C(═NR)—NH-8-C(—NHR)═N—) tautomers. The latter two areparticularly common in heteroaryl and heterocyclic rings and the presentinvention encompasses all tautomeric forms of the compounds.

Technical and scientific terms used herein have the meaning commonlyunderstood by one of skill in the art to which the present inventionpertains, unless otherwise defined. Reference is made herein to variousmethodologies and materials known to those of skill in the art. Standardreference works setting forth the general principles of pharmacologyinclude Goodman and Gilman's The Pharmacological Basis of Therapeutics,10^(th) Ed., McGraw Hill Companies Inc., New York (2001). Any suitablematerials and/or methods known to those of skill can be utilized incarrying out the present invention. However, preferred materials andmethods are described. Materials, reagents and the like to whichreference are made in the following description and examples areobtainable from commercial sources, unless otherwise noted.

The term “acyl” as used herein denotes a group of formula —C(═O)Rwherein R is hydrogen or lower alkyl as defined herein. The term or“alkylcarbonyl” as used herein denotes a group of formula C(═O)R whereinR is alkyl as defined herein. The term C₁₋₆ acyl refers to a group—C(═O)R contain 6 carbon atoms. The term “arylcarbonyl” as used hereinmeans a group of formula C(═O)R wherein R is an aryl group; the term“benzoyl” as used herein an “arylcarbonyl” group wherein R is phenyl.

The term “alkyl” as used herein denotes an unbranched or branched chain,saturated, monovalent hydrocarbon residue containing 1 to 10 carbonatoms. The term “lower alkyl” denotes a straight or branched chainhydrocarbon residue containing 1 to 6 carbon atoms. “C₁₋₁₀ alkyl” asused herein refers to an alkyl composed of 1 to 10 carbons. Examples ofalkyl groups include, but are not limited to, lower alkyl groups includemethyl, ethyl, propyl, i-propyl, n-butyl, i-butyl, t-butyl or pentyl,isopentyl, neopentyl, hexyl, heptyl, and octyl.

When the term “alkyl” is used as a suffix following another term, as in“phenylalkyl,” or “hydroxyalkyl,” this is intended to refer to an alkylgroup, as defined above, being substituted with one to two substituentsselected from the other specifically-named group. Thus, for example,“phenylalkyl” denotes the radical R′R″-, wherein R′ is a phenyl radical,and R″ is an alkylene radical as defined herein with the understandingthat the attachment point of the phenylalkyl moiety will be on thealkylene radical. Examples of arylalkyl radicals include, but are notlimited to, benzyl, phenylethyl, 3-phenylpropyl. The terms “arylalkyl”,“aryl alkyl”, or “aralkyl” are interpreted similarly except R′ is anaryl radical. The terms “heteroaryl alkyl” or “heteroarylalkyl” areinterpreted similarly except R′ is optionally an aryl or a heteroarylradical.

The term “haloalkyl” as used herein denotes a unbranched or branchedchain alkyl group as defined above wherein 1, 2, 3 or more hydrogenatoms are substituted by a halogen. The term “lower haloalkyl” denotes astraight or branched chain hydrocarbon residue containing 1 to 6 carbonatoms, wherein 1, 2, 3 or more hydrogen atoms are substituted by ahalogen. Examples are 1-fluoromethyl, 1-chloromethyl, 1-bromomethyl,1-iodomethyl, difluoromethyl, trifluoromethyl, trichloromethyl,tribromomethyl, triiodomethyl, 1-fluoroethyl, 1-chloroethyl,1-bromoethyl, 1-iodoethyl, 2-fluoroethyl, 2-chloroethyl, 2-bromoethyl,2-iodoethyl, 2,2-dichloroethyl, 3-bromopropyl or 2,2,2-trifluoroethyl.

The term “alkylene” as used herein denotes a divalent saturated linearhydrocarbon radical of 1 to 10 carbon atoms (e.g., (CH₂)_(n)) or abranched saturated divalent hydrocarbon radical of 2 to 10 carbon atoms(e.g., —CHMe- or —CH₂CH(i-Pr)CH₂—), unless otherwise indicated. Exceptin the case of methylene, the open valences of an alkylene group are notattached to the same atom. Examples of alkylene radicals include, butare not limited to, methylene, ethylene, propylene, 2-methyl-propylene,1,1-dimethyl-ethylene, butylene, 2-ethylbutylene.

The term “alkoxy” as used herein means an —O-alkyl group, wherein alkylis as defined above such as methoxy, ethoxy, n-propyloxy, i-propyloxy,n-butyloxy, i-butyloxy, t-butyloxy, pentyloxy, hexyloxy, including theirisomers. “Lower alkoxy” as used herein denotes an alkoxy group with a“lower alkyl” group as previously defined. “C₁₋₁₀ alkoxy” as used hereinrefers to an —O-alkyl wherein alkyl is C₁₋₁₀.

The term “hydroxyalkyl” as used herein denotes an alkyl radical asherein defined wherein one to three hydrogen atoms on different carbonatoms is/are replaced by hydroxyl groups.

The term “cycloalkyl” as used herein refers to a saturated carbocyclicring containing 3 to 8 carbon atoms, i.e. cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cycloheptyl or cyclooctyl. “C₃₋₇ cycloalkyl” asused herein refers to an cycloalkyl composed of 3 to 7 carbons in thecarbocyclic ring.

The term “halogen” or “halo” as used herein means fluorine, chlorine,bromine, or iodine.

The term “heteroaryl” or “heteroaromatic” as used herein means amonocyclic or bicyclic, or tricyclic radical of 5 to 18 ring atomshaving at least one aromatic ring containing four to eight atoms perring, incorporating one or more N, O, or S heteroatoms, the remainingring atoms being carbon, with the understanding that the attachmentpoint of the heteroaryl radical will be on an aromatic ring. As wellknown to those skilled in the art, heteroaryl rings have less aromaticcharacter than their all-carbon counter parts. Thus, for the purposes ofthe invention, a heteroaryl group need only have some degree of aromaticcharacter. Examples of heteroaryl moieties include monocyclic aromaticheterocycles having 5 to 6 ring atoms and 1 to 3 heteroatoms include,but is not limited to, pyridinyl, pyrimidinyl, pyrazinyl, pyrrolyl,pyrazolyl, imidazolyl, oxazol, isoxazole, thiazole, isothiazole,triazoline, thiadiazole and oxadiaxoline which can optionally besubstituted with one or more, preferably one or two substituentsselected from hydroxy, cyano, alkyl, alkoxy, thio, lower haloalkoxy,alkylthio, halo, haloalkyl, alkylsulfinyl, alkylsulfonyl, halogen,amino, alkylamino,dialkylamino, aminoalkyl, alkylaminoalkyl, anddialkylaminoalkyl, nitro, alkoxycarbonyl and carbamoyl, alkylcarbamoyl,dialkylcarbamoyl, arylcarbamoyl, alkylcarbonylamino andarylcarbonylamino. Examples of bicyclic moieties include, but are notlimited to, quinolinyl, isoquinolinyl, benzofuryl, benzothiophenyl,benzoxazole, benzisoxazole, benzothiazole and benzisothiazole.

The term “heterocycloalkyl”, “heterocyclyl” or “heterocycle” as usedherein denotes a monovalent saturated cyclic radical, consisting of oneor more rings, preferably one to two rings, or three rings, of three toeight atoms per ring, incorporating one or more ring carbon atoms andone or more ring heteroatoms (chosen from N,O or S(═O)₀₋₂), wherein thepoint of attachment can be through either a carbon atom or a heteroatom,and which can optionally be independently substituted with one or more,preferably one or two or three substituents selected from hydroxy, oxo,cyano, lower alkyl, lower alkoxy, lower haloalkoxy, alkylthio, halo,haloalkyl, hydroxyalkyl, nitro, alkoxycarbonyl, amino, alkylamino,alkylsulfonyl, arylsulfonyl, alkylaminosulfonyl, arylaminosulfonyl,alkylsulfonylamino, arylsulfonylamino, alkylaminocarbonyl,arylaminocarbonyl, alkylcarbonylamino, arylcarbonylamino, unlessotherwise indicated. Examples of heterocyclic radicals include, but arenot limited to, azetidinyl, pyrrolidinyl, hexahydroazepinyl, oxetanyl,tetrahydrofuranyl, tetrahydrothiophenyl, oxazolidinyl, thiazolidinyl,isoxazolidinyl, morpholinyl, piperazinyl, piperidinyl,tetrahydropyranyl, thiomorpholinyl, quinuclidinyl and imidazolinyl.

The phrase “organ rejection” includes acute allograft or xenograftrejection and chronic allograft or xenograft rejection in the setting ofvascularized and/or non-vascularized (e.g. bone marrow, pancreatic isletcells) transplants.

Commonly used abbreviations include: acetyl (Ac),azo-bis-isobutyrylnitrile (AIBN), atmospheres (Atm),9-borabicyclo[3.3.1]nonane (9-BBN or BBN), tert-butoxycarbonyl (Boc),di-tert-butyl pyrocarbonate or boc anhydride (BOC₂O), benzyl (Bn), butyl(Bu), Chemical Abstracts Registration Number (CASRN), benzyloxycarbonyl(CBZ or Z), carbonyl diimidazole (CDI), 1,4-diazabicyclo[2.2.2]octane(DABCO), diethylaminosulfur trifluoride (DAST), dibenzylideneacetone(dba), 1,5-diazabicyclo[4.3.0]non-5-ene (DBN),1,8-diazabicyclo[5.4.0]undec-7-ene (DBU), N,N′-dicyclohexylcarbodiimide(DCC), 1,2-dichloroethane (DCE), dichloromethane (DCM), diethylazodicarboxylate (DEAD), di-iso-propylazodicarboxylate (DIAD),di-iso-butylaluminumhydride (DIBAL or DIBAL-H), di-iso-propylethylamine(DIPEA), N,N-dimethyl acetamide (DMA), 4-N,N-dimethylaminopyridine(DMAP), N,N-dimethylformamide (DMF), dimethyl sulfoxide (DMSO),1,1′-bis-(diphenylphosphino)ethane (dppe),1,1′-bis-(diphenylphosphino)ferrocene (dppf),1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI),ethyl (Et), ethyl acetate (EtOAc), ethanol (EtOH),2-ethoxy-2H-quinoline-1-carboxylic acid ethyl ester (EEDQ), diethylether (Et₂O), O-(7-azabenzotriazole-1-yl)-N,N,N′N′-tetramethyluroniumhexafluorophosphate acetic acid (HATU), acetic acid (HOAc),1-N-hydroxybenzotriazole (HOBt), high pressure liquid chromatography(HPLC), iso-propanol (IPA), lithium hexamethyl disilazane (LiHMDS),methanol (MeOH), melting point (mp), MeSO₂— (mesyl or Ms), methyl (Me),acetonitrile (MeCN), m-chloroperbenzoic acid (MCPBA), mass spectrum(ms), methyl t-butyl ether (MTBE), N-bromosuccinimide (NBS),N-carboxyanhydride (NCA), N-chlorosuccinimide (NCS), N-methylmorpholine(NMM), N-methylpyrrolidone (NMP), pyridinium chlorochromate (PCC),pyridinium dichromate (PDC), phenyl (Ph), propyl (Pr), iso-propyl(i-Pr), pounds per square inch (psi), pyridine (pyr), room temperature(rt or RT), tert-butyldimethylsilyl or t-BuMe₂Si (TBDMS), triethylamine(TEA or Et₃N), 2,2,6,6-tetramethylpiperidine 1-oxyl (TEMPO), triflate orCF₃SO₂— (Tf), trifluoroacetic acid (TFA),1,1′-bis-2,2,6,6-tetramethylheptane-2,6-dione (TMHD),O-benzotriazol-1-yl-N,N,N′,N′-tetramethyluronium tetrafluoroborate(TBTU), thin layer chromatography (TLC), tetrahydrofuran (THF),trimethylsilyl or Me₃Si (TMS), p-toluenesulfonic acid monohydrate (TsOHor pTsOH), 4-Me-C₆H₄SO₂— or tosyl (Ts), N-urethane-N-carboxyanhydride(UNCA). Conventional nomenclature including the prefixes normal (n), iso(i-), secondary (sec-), tertiary (tert-) and neo have their customarymeaning when used with an alkyl moiety. (J. Rigaudy and D. P. Klesney,Nomenclature in Organic Chemistry, IUPAC 1979 Pergamon Press, Oxford).

Compounds and Preparation

Examples of representative compounds encompassed by the presentinvention and within the scope of the invention are provided in thefollowing Table. These examples and preparations which follow areprovided to enable those skilled in the art to more clearly understandand to practice the present invention. They should not be considered aslimiting the scope of the invention, but merely as being illustrativeand representative thereof.

In general, the nomenclature used in this Application is based onAUTONOM™ v.4.0, a Beilstein Institute computerized system for thegeneration of IUPAC systematic nomenclature. If there is a discrepancybetween a depicted structure and a name given that structure, thedepicted structure is to be accorded more weight. In addition, if thestereochemistry of a structure or a portion of a structure is notindicated with, for example, bold or dashed lines, the structure orportion of the structure is to be interpreted as encompassing allstereoisomers of it.

TABLE I depicts exemplified compounds according to Formula I.

TABLE I SYSTEMATIC COMPOUND NAME STRUCTURE MP I-1 2-(Cyclopentyl-methyl-amino)-5H- pyrrolo[2,3- b]pyrazine- carboxylic acidisopropylamide

277-278 I-2 1-[7-(2,2- Dimethyl- propionyl)-5H- pyrrolo[2,3-b]pyrazin-2-yl]-2,2- dimethyl-propan-1- one

188-198 I-3 1-(2- Cyclopentyloxy- 5H-pyrrolo[2,3- b]pyrazin-7-yl)-2,2-dimethyl-propan-1- one

I-4 N-[7-(2,2- Dimethyl- propionyl)-5H- pyrrolo[2,3- b]pyrazin-2-yl]-benzamide

I-5 Cyclo- hexanecarboxylic acid [7-(2,2- dimethyl- propionyl)-5H-pyrrolo[2,3- b]pyrazin-2-yl]- amide

I-6 N-[7-(2,2- Dimethyl- propionyl)-5H- pyrrolo[2,3- b]pyrazin-2-yl]-N-methyl-benzamide

I-7 N-[7-(2,2- Dimethyl- propionyl)-5H- pyrrolo[2,3- b]pyrazin-2-yl]-benzenesulfonamide

I-8 N-[7-(2,2- Dimethyl- propionyl)-5H- pyrrolo[2,3- b]pyrazin-2-yl]-N-methyl- benzenesulfonamide

I-9 7-(2,2-Dimethyl- propionyl)-5H- pyrrolo[2,3- b]pyrazine-2-carboxylic acid dimethylamide

I-10 7-(2,2-Dimethyl- propionyl)-5H- pyrrolo[2,3- b]pyrazin-2-carboxylic acid isopropylamide

I-11 1-(2-Acetyl-5H- pyrrolo[2,3- b]pyrazin-7-yl)-2,2-dimethyl-propan-1- one

I-12 1-(2-Isobutyryl 5H-pyrrolo[2,3- b]pyrazin-7-yl)-2,2-dimethyl-propan-1- one

I-13 2,2-Dimethyl-1-[2- (2-methyl- benzoyl)-5H- pyrrolo[2,3-b]pyrazin-7-yl]- propan-1-one

I-14 1-(2-Benzoyl-5H- pyrrolo[2,3- b]pyrazin-7-yl)-2,2-dimethyl-propan-1- one

I-15 7-(2,2-Dimethyl- propionyl)-5H- pyrrolo[2,3- b]pyrazin-2-carboxylic acid cyclopentylamide

I-16 2,2-Dimethyl-1-[2- (pyridine-4- carbonyl)-5H- pyrrolo[2,3-b]pyrazin-7-yl]- propan-1-one

I-17 2,2-Dimethyl-1-[2- (pyridine-3- carbonyl)-5H- pyrrolo[2,3-b]pyrazin-7-yl]- propan-1-one

I-18 1-(2- Cycloheptyloxy- 5H-pyrrolo[2,3- b]pyrazin-7-yl)-2,2-dimethyl-propan-1- one

I-19 1-(2- Cyclohexyloxy- 5H-pyrrolo[2,3- b]pyrazin-7-yl)-2,2-dimethyl-propan-1- one

I-20 1-(2-Isopropoxy- 5H-pyrrolo[2,3- b]pyrazin-7-yl)-2,2-dimethyl-propan-1- one

I-21 1-(2-Ethoxy- 5H-pyrrolo[2,3- b]pyrazin-7-yl)-2,2-dimethyl-propan-1- one

I-22 2,2-Dimethyl-1-[2- (2-morpholin-4-yl- 2-oxo-ethoxy)-5H-pyrrolo[2,3- b]pyrazin-7-yl]- propan-1-one

I-23 1-[2-(1,2- Dimethyl- propoxy)-5H- pyrrolo[2,3- b]pyrazin-7-yl]-2,2-dimethyl-propan-1- one

I-24 1-(2-Isobutoxy- 5H-pyrrolo[2,3- b]pyrazin-7-yl)-2,2-dimethyl-propan-1- one

I-25 1-[2-(1-Benzyl- 1H-[1,2,3]triazol-4- yl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2- dimethyl-propan-1- one

I-26 1-Cyclohexyl-3-[7- (2,2-dimethyl- propionyl)-5H- pyrrolo[2,3-b]pyrazin-2-yl]- urea

I-27 1-Cycloheptyl-3-[7- (2,2-dimethyl- propionyl)-5H- pyrrolo[2,3-b]pyrazin-2-yl]- urea

I-28 2,2-Dimethyl-1-[2- (methyl-phenyl- amino)-5H- pyrrolo[2,3-b]pyrazin-7-yl]- propan-1-one

I-29 1-(2-sec-Butoxy- 5H-pyrrolo[2,3- b]pyrazin-7-yl)-2,2-dimethyl-propan-1- one

I-30 2,2-Dimethyl-1-(2- phenylamino-5H- pyrrolo[2,3- b]pyrazin-7-yl)-propan-1-one

I-31 1-(2- Cyclopentyloxy- 5H-pyrrolo[2,3- b]pyrazin-7-yl)-2,2-dimethyl-propan-1- one

I-32 1-[2-(Cyclopentyl- methyl-amino)-5H- pyrrolo[2,3-b]pyrazin-7-yl]-2,2- dimethyl-propan-1- one

I-33 2,2-Dimethyl-1-(2- phenoxy-5H- pyrrolo[2,3- b]pyrazin-7-yl)-propan-1-one

I-34 2,2-Dimethyl-1-{2- [4-(4-methyl- piperazin-1-yl)- phenylamino]-5H-pyrrolo[2,3- b]pyrazin-7-yl}- propan-1-one

I-35 2,2-Dimethyl-1-{2- [3-(4-methyl- piperazin-1-yl)- phenylamino]-5H-pyrrolo[2,3- b]pyrazin-7-yl}- propan-1-one

I-36 1-Cyclohexyl-3-[7- (2,2-dimethyl- propionyl)-5H- pyrrolo[2,3-b]pyrazin-2-yl]- urea

I-37 1-Cycloheptyl-3-[7- (2,2-dimethyl- propionyl)-5H- pyrrolo[2,3-b]pyrazin-2-yl]- urea

I-38 1-[7-(2,2- Dimethyl- propionyl)-5H- pyrrolo[2,3-b]pyrazin-2-yl]-2,2- dimethyl-propan-1- one

I-39 1-[2-(6-Methoxy- pyridin-2-ylamino)- 5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2- dimethyl-propan-1- one

Dosage and Administration

The compounds of the present invention may be formulated in a widevariety of oral administration dosage forms and carriers. Oraladministration can be in the form of tablets, coated tablets, dragees,hard and soft gelatine capsules, solutions, emulsions, syrups, orsuspensions. Compounds of the present invention are efficacious whenadministered by other routes of administration including continuous(intravenous drip) topical parenteral, intramuscular, intravenous,subcutaneous, transdermal (which may include a penetration enhancementagent), buccal, nasal, inhalation and suppository administration, amongother routes of administration. The preferred manner of administrationis generally oral using a convenient daily dosing regimen which can beadjusted according to the degree of affliction and the patient'sresponse to the active ingredient.

A compound or compounds of the present invention, as well as theirpharmaceutically useable salts, together with one or more conventionalexcipients, carriers, or diluents, may be placed into the form ofpharmaceutical compositions and unit dosages. The pharmaceuticalcompositions and unit dosage forms may be comprised of conventionalingredients in conventional proportions, with or without additionalactive compounds or principles, and the unit dosage forms may containany suitable effective amount of the active ingredient commensurate withthe intended daily dosage range to be employed. The pharmaceuticalcompositions may be employed as solids, such as tablets or filledcapsules, semisolids, powders, sustained release formulations, orliquids such as solutions, suspensions, emulsions, elixirs, or filledcapsules for oral use; or in the form of suppositories for rectal orvaginal administration; or in the form of sterile injectable solutionsfor parenteral use. A typical preparation will contain from about 5% toabout 95% active compound or compounds (w/w). The term “preparation” or“dosage form” is intended to include both solid and liquid formulationsof the active compound and one skilled in the art will appreciate thatan active ingredient can exist in different preparations depending onthe target organ or tissue and on the desired dose and pharmacokineticparameters.

The term “excipient” as used herein refers to a compound that is usefulin preparing a pharmaceutical composition, generally safe, non-toxic andneither biologically nor otherwise undesirable, and includes excipientsthat are acceptable for veterinary use as well as human pharmaceuticaluse. The compounds of this invention can be administered alone but willgenerally be administered in admixture with one or more suitablepharmaceutical excipients, diluents or carriers selected with regard tothe intended route of administration and standard pharmaceuticalpractice.

“Pharmaceutically acceptable” means that which is useful in preparing apharmaceutical composition that is generally safe, non-toxic, andneither biologically nor otherwise undesirable and includes that whichis acceptable for veterinary as well as human pharmaceutical use.

A “pharmaceutically acceptable salt” form of an active ingredient mayalso initially confer a desirable pharmacokinetic property on the activeingredient which were absent in the non-salt form, and may evenpositively affect the pharmacodynamics of the active ingredient withrespect to its therapeutic activity in the body. The phrase“pharmaceutically acceptable salt” of a compound means a salt that ispharmaceutically acceptable and that possesses the desiredpharmacological activity of the parent compound. Such salts include: (1)acid addition salts, formed with inorganic acids such as hydrochloricacid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, andthe like; or formed with organic acids such as acetic acid, propionicacid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvicacid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid,fumaric acid, tartaric acid, citric acid, benzoic acid,3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid,2-hydroxyethanesulfonic acid, benzenesulfonic acid,4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid,4-toluenesulfonic acid, camphorsulfonic acid,4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid,3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid,lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoicacid, salicylic acid, stearic acid, muconic acid, and the like; or (2)salts formed when an acidic proton present in the parent compound eitheris replaced by a metal ion, e.g., an alkali metal ion, an alkaline earthion, or an aluminum ion; or coordinates with an organic base such asethanolamine, diethanolamine, triethanolamine, tromethamine,N-methylglucamine, and the like.

Solid form preparations include powders, tablets, pills, capsules,cachets, suppositories, and dispersible granules. A solid carrier may beone or more substances which may also act as diluents, flavoring agents,solubilizers, lubricants, suspending agents, binders, preservatives,tablet disintegrating agents, or an encapsulating material. In powders,the carrier generally is a finely divided solid which is a mixture withthe finely divided active component. In tablets, the active componentgenerally is mixed with the carrier having the necessary bindingcapacity in suitable proportions and compacted in the shape and sizedesired. Suitable carriers include but are not limited to magnesiumcarbonate, magnesium stearate, talc, sugar, lactose, pectin, dextrin,starch, gelatin, tragacanth, methylcellulose, sodiumcarboxymethylcellulose, a low melting wax, cocoa butter, and the like.Solid form preparations may contain, in addition to the activecomponent, colorants, flavors, stabilizers, buffers, artificial andnatural sweeteners, dispersants, thickeners, solubilizing agents, andthe like.

Liquid formulations also are suitable for oral administration includeliquid formulation including emulsions, syrups, elixirs, aqueoussolutions, aqueous suspensions. These include solid form preparationswhich are intended to be converted to liquid form preparations shortlybefore use. Emulsions may be prepared in solutions, for example, inaqueous propylene glycol solutions or may contain emulsifying agentssuch as lecithin, sorbitan monooleate, or acacia. Aqueous solutions canbe prepared by dissolving the active component in water and addingsuitable colorants, flavors, stabilizing, and thickening agents. Aqueoussuspensions can be prepared by dispersing the finely divided activecomponent in water with viscous material, such as natural or syntheticgums, resins, methylcellulose, sodium carboxymethylcellulose, and otherwell known suspending agents.

The compounds of the present invention may be formulated for parenteraladministration (e.g., by injection, for example bolus injection orcontinuous infusion) and may be presented in unit dose form in ampoules,pre-filled syringes, small volume infusion or in multi-dose containerswith an added preservative. The compositions may take such forms assuspensions, solutions, or emulsions in oily or aqueous vehicles, forexample solutions in aqueous polyethylene glycol. Examples of oily ornonaqueous carriers, diluents, solvents or vehicles include propyleneglycol, polyethylene glycol, vegetable oils (e.g., olive oil), andinjectable organic esters (e.g., ethyl oleate), and may containformulatory agents such as preserving, wetting, emulsifying orsuspending, stabilizing and/or dispersing agents. Alternatively, theactive ingredient may be in powder form, obtained by aseptic isolationof sterile solid or by lyophilisation from solution for constitutionbefore use with a suitable vehicle, e.g., sterile, pyrogen-free water.

The compounds of the present invention may be formulated for topicaladministration to the epidermis as ointments, creams or lotions, or as atransdermal patch. Ointments and creams may, for example, be formulatedwith an aqueous or oily base with the addition of suitable thickeningand/or gelling agents. Lotions may be formulated with an aqueous or oilybase and will in general also containing one or more emulsifying agents,stabilizing agents, dispersing agents, suspending agents, thickeningagents, or coloring agents. Formulations suitable for topicaladministration in the mouth include lozenges comprising active agents ina flavored base, usually sucrose and acacia or tragacanth; pastillescomprising the active ingredient in an inert base such as gelatin andglycerin or sucrose and acacia; and mouthwashes comprising the activeingredient in a suitable liquid carrier.

The compounds of the present invention may be formulated foradministration as suppositories. A low melting wax, such as a mixture offatty acid glycerides or cocoa butter is first melted and the activecomponent is dispersed homogeneously, for example, by stirring. Themolten homogeneous mixture is then poured into convenient sized molds,allowed to cool, and to solidify.

The compounds of the present invention may be formulated for vaginaladministration. Pessaries, tampons, creams, gels, pastes, foams orsprays containing in addition to the active ingredient such carriers asare known in the art to be appropriate.

The compounds of the present invention may be formulated for nasaladministration. The solutions or suspensions are applied directly to thenasal cavity by conventional means, for example, with a dropper, pipetteor spray. The formulations may be provided in a single or multidoseform. In the latter case of a dropper or pipette, this may be achievedby the patient administering an appropriate, predetermined volume of thesolution or suspension. In the case of a spray, this may be achieved forexample by means of a metering atomizing spray pump.

The compounds of the present invention may be formulated for aerosoladministration, particularly to the respiratory tract and includingintranasal administration. The compound will generally have a smallparticle size for example of the order of five (5) microns or less. Sucha particle size may be obtained by means known in the art, for exampleby micronization. The active ingredient is provided in a pressurizedpack with a suitable propellant such as a chlorofluorocarbon (CFC), forexample, dichlorodifluoromethane, trichlorofluoromethane, ordichlorotetrafluoroethane, or carbon dioxide or other suitable gas. Theaerosol may conveniently also contain a surfactant such as lecithin. Thedose of drug may be controlled by a metered valve. Alternatively theactive ingredients may be provided in a form of a dry powder, forexample a powder mix of the compound in a suitable powder base such aslactose, starch, starch derivatives such as hydroxypropylmethylcellulose and polyvinylpyrrolidine (PVP). The powder carrier will form agel in the nasal cavity. The powder composition may be presented in unitdose form for example in capsules or cartridges of e.g., gelatin orblister packs from which the powder may be administered by means of aninhaler.

When desired, formulations can be prepared with enteric coatings adaptedfor sustained or controlled release administration of the activeingredient. For example, the compounds of the present invention can beformulated in transdermal or subcutaneous drug delivery devices. Thesedelivery systems are advantageous when sustained release of the compoundis necessary and when patient compliance with a treatment regimen iscrucial. Compounds in transdermal delivery systems are frequentlyattached to an skin-adhesive solid support. The compound of interest canalso be combined with a penetration enhancer, e.g., Azone(1-dodecylaza-cycloheptan-2-one). Sustained release delivery systems areinserted subcutaneously into to the subdermal layer by surgery orinjection. The subdermal implants encapsulate the compound in a lipidsoluble membrane, e.g., silicone rubber, or a biodegradable polymer,e.g., polyactic acid.

Suitable formulations along with pharmaceutical carriers, diluents andexcipients are described in Remington: The Science and Practice ofPharmacy 1995, edited by E. W. Martin, Mack Publishing Company, 19thedition, Easton, Pa. A skilled formulation scientist may modify theformulations within the teachings of the specification to providenumerous formulations for a particular route of administration withoutrendering the compositions of the present invention unstable orcompromising their therapeutic activity.

The modification of the present compounds to render them more soluble inwater or other vehicle, for example, may be easily accomplished by minormodifications (salt formulation, esterification, etc.), which are wellwithin the ordinary skill in the art. It is also well within theordinary skill of the art to modify the route of administration anddosage regimen of a particular compound in order to manage thepharmacokinetics of the present compounds for maximum beneficial effectin patients.

The term “therapeutically effective amount” as used herein means anamount required to reduce symptoms of the disease in an individual. Thedose will be adjusted to the individual requirements in each particularcase. That dosage can vary within wide limits depending upon numerousfactors such as the severity of the disease to be treated, the age andgeneral health condition of the patient, other medicaments with whichthe patient is being treated, the route and form of administration andthe preferences and experience of the medical practitioner involved. Fororal administration, a daily dosage of between about 0.01 and about 1000mg/kg body weight per day should be appropriate in monotherapy and/or incombination therapy. A preferred daily dosage is between about 0.1 andabout 500 mg/kg body weight, more preferred 0.1 and about 100 mg/kg bodyweight and most preferred 1.0 and about 10 mg/kg body weight per day.Thus, for administration to a 70 kg person, the dosage range would beabout 7 mg to 0.7 g per day. The daily dosage can be administered as asingle dosage or in divided dosages, typically between 1 and 5 dosagesper day. Generally, treatment is initiated with smaller dosages whichare less than the optimum dose of the compound. Thereafter, the dosageis increased by small increments until the optimum effect for theindividual patient is reached. One of ordinary skill in treatingdiseases described herein will be able, without undue experimentationand in reliance on personal knowledge, experience and the disclosures ofthis application, to ascertain a therapeutically effective amount of thecompounds of the present invention for a given disease and patient.

The pharmaceutical preparations are preferably in unit dosage forms. Insuch form, the preparation is subdivided into unit doses containingappropriate quantities of the active component. The unit dosage form canbe a packaged preparation, the package containing discrete quantities ofpreparation, such as packeted tablets, capsules, and powders in vials orampoules. Also, the unit dosage form can be a capsule, tablet, cachet,or lozenge itself, or it can be the appropriate number of any of thesein packaged form.

The following examples illustrate the preparation and biologicalevaluation of compounds within the scope of the invention. Theseexamples and preparations which follow are provided to enable thoseskilled in the art to more clearly understand and to practice thepresent invention. They should not be considered as limiting the scopeof the invention, but merely as being illustrative and representativethereof.

EXAMPLES Example 1

1-[2-bromo-5-(2-trimethylsilanyl-ethoxymethyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-one

Sodium hydride (60% in mineral oil, 0.019 g, 0.48 mmol) was added to astirring solution of1-(2-bromo-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one(0.094 g, 0.33 mmol) in 1.5 mL of N,N′-dimethylformamide at 0-5° C. Thebubbling yellow mixture was stirred at 0-5° C. for 15 min., then2-(trimethylsilyl)ethoxymethyl chloride (0.075 mL, 0.42 mmol) was added.The resulting cloudy yellow mixture was stirred at RT for 3 h, thenpartitioned between 10 mL of water and 10 mL of ethyl acetate. Theorganic layer was sequentially washed with two 10 mL portions of waterand 10 mL of a sat. aq. NaCl solution, dried over MgSO₄, filtered, andconcentrated to an orange oil. Silica gel chromatography (10%EtOAc/hexanes) afforded 0.129 g (93%) of slightly impure1-[2-bromo-5-(2-trimethylsilanyl-ethoxymethyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-oneas a yellow oil that was used without further purification.

Example 2

7-(2,2-dimethyl-propionyl)-5-(2-trimethylsilanyl-ethoxymethyl)-1,5-dihydro-pyrrolo[2,3-b]pyrazin-2-one

A mixture of1-[2-bromo-5-(2-trimethylsilanyl-ethoxymethyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-one(0.067 g, 0.16 mmol), 0.4 mL dioxane, 0.4 mL water, Pd₂(dba)₃ (0.009 g,0.01 mmol), 2-di-t-butylphosphino-2′,4′,6′-triisopropyl-1,1′-biphenyl(0.008 g, 0.02 mmol), and freshly ground KOH (0.038 g, 0.67 mmol) in asealed tube under N₂ was stirred at 100° C. for 15 h. The resultingorange-black mixture was partitioned between 5 mL of ethyl acetate and 5mL of water, with a few drops of ethanol added to reduce emulsions. Theaqueous layer was extracted with 5 mL of ethyl acetate. The combinedorganic layers were dried over Na₂SO₄, filtered, and concentrated to anorange oil. Silica gel chromatography (20→40% EtOAc/hexanes) afforded0.029 g (51%) of7-(2,2-dimethyl-propionyl)-5-(2-trimethylsilanyl-ethoxymethyl)-1,5-dihydro-pyrrolo[2,3-b]pyrazin-2-oneas a pale yellow solid.

Example 3

1-[2-cyclopentyloxy-5-(2-trimethylsilanyl-ethoxymethyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-one

A solution of7-(2,2-dimethyl-propionyl)-5-(2-trimethylsilanyl-ethoxymethyl)-1,5-dihydro-pyrrolo[2,3-b]pyrazin-2-one(0.029 g, 0.082 mmol), 1 mL of tetrahydrofuran, cyclopentanol (0.010 mL,0.11 mmol), tributylphosphine (0.025 mL, 0.1 mmol), and diisopropylazodicarboxylate (0.020 mL, 0.10 mmol) under nitrogen was stirred at 65°C. for 17.5 h then allowed to cool. Additional cyclopentanol (0.020 mL,0.22 mmol), tributylphosphine (0.050 mL, 0.20 mmol), and diisopropylazodicarboxylate (0.040 mL, 0.21 mmol) were added, and the solutionstirred at 65° C. for 2.5 h. The yellow solution was partitioned between5 mL of water and 5 mL of ethyl acetate. The organic layer was washedwith 5 mL of a sat. aq. NaCl solution, dried over MgSO₄, filtered andconcentrated to a yellow oil. Silica gel chromatography (10%EtOAc/hexanes) afforded 0.017 g (51%) of1-[2-cyclopentyloxy-5-(2-trimethylsilanyl-ethoxymethyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-oneas a colorless oil.

Example 4

1-(2-cyclopentyloxy-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one

A solution of1-[2-cyclopentyloxy-5-(2-trimethylsilanyl-ethoxymethyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-one(0.017 g, 0.041 mmol) in 1 mL of dichloromethane and 1 mL oftrifluoroacetic acid was stirred for 5 h then concentrated to a yellowoil. Silica gel chromatography (0→40% EtOAc/hexanes) afforded 0.012 g ofa white solid. ¹H NMR spectroscopy indicated that this intermediate wasthe formaldehyde adduct of the desired product. The solid was dissolvedin 1 mL of ethanol and the solution was treated with sodium acetatetrihydrate (0.064 g, 0.47 mmol). The colorless mixture was stirred for 5h then concentrated. The residue was partitioned between 5 mL of ethylacetate and 5 mL of water. The organic layer was sequentially washedwith 5 mL of water and 5 mL of a sat. aq. NaCl solution, dried overMgSO₄, filtered and concentrated to 0.007 g (57%) of1-(2-cyclopentyloxy-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-oneas a slightly impure white solid.

The following compounds were prepared according to the above generalprocedure:

1-(2-Cycloheptyloxy-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one

1-(2-Cyclohexyloxy-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one

1-(2-Isopropoxy-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one

1-[2-(1,2-Dimethyl-propoxy)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-one

1-(2-Isobutoxy-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one

1-(2-sec-Butoxy-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one

Example 5

2-(Cyclopentyl-methyl-amino)-5H-pyrrolo[2,3-b]pyrazine-7-carboxylic acidisopropylamide

Step 1. Pyrrolopyrazine C (340 mg, 1.7 mmol) andN-methyl-cyclopentylamine were dissolved in NMP (2 mL) and placed in asealed tube. The solution was irradiated with high intensity microwaveat 240 Celsius for 1.5 hours. Upon cooling the mixture was partitionedbetween water and ethyl acetate. The extract was aged over anhydroussodium sulfate and the volatiles were removed. The desired adduct (A,102 mg) was isolated by SGC eluted with 10 to 60% ethyl acetate inhexanes and displayed spectroscopic properties consistent with theproposed structure.

Step 2. Pyrrolopyrazine A (100 mg, 0.46 mmol) was dissolved inN,N-dimethylformamide (5 mL) and cooled to 0 Celsius. Powdered potassiumhydroxide (67 mg, 1.2 mmol) and iodine (127 mg, 0.5 mmol) were added insequence which gave a tan solution after 30 minutes at 0 Celsius. Thesolution was treated with water, decolorized with 10% sodiumthiosulfate_((aq)) and the resulting precipitate was filtered. Thefilter cake was washed with fresh water and stored in vacuo at 50Celsius overnight. The tan powder (B, 120 mg) displayed spectroscopicproperties consistent with the proposed structure.

Step 3. The desired amide I was prepared according to step 2 with B (110mg, 0.32 mmol) and K₂CO₃ (135 mg, 1.0 mmol) by replacing the amine withiso-propylamine (0.08 mL, 1.0 mmol). The amide I was obtained as a solid(35 mg): mp 277-278° C.; ESMS m/z 302 (M⁺¹) for MW of 301.

2-(3-Methylamino-piperidin-1-yl)-5H-pyrrolo[2,3-b]pyrazine-7-carboxylicacid isopropylamide was obtained by replacing N-methylcyclopentyl aminewith 3-methylaminopiperidine in step 1 and conducting the reaction for 4hours. The crude reaction mixture treated with di-tert-butyl dicarbonateand 4-dimethylaminopyridine in reluxing tetrahydrofuran and subjected tosilica gel chromatography (eluant: 20 to 80% ethyl acetate in hexanes.The title compound was obtained by replacing B in step 2 with[1-(7-Iodo-5H-pyrrolo[2,3-b]pyrazin-2-yl)-piperidin-3-yl]-methyl-carbamicacid tert-butyl ester then the product of the carbonylation was treatedwith trifluoroacetic acid in refluxing 1,2-dichloroethane to obtain thetitle compound as the trifluoroacetate salt: m.p. 54-64° C.; MS m/z 317(M^(+H) for free base).

Example 6

7-(2,2-Dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazine-2-carboxylic aciddimethylamide:1-(2-bromo-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one (50mg, 0.177 mmol) was dissolved in N,N-dimethylformamide and the flask waspurged with carbon monoxide. Dimethylamine (0.32 ml of a 5.5M ethanolsolution) was added followed by palladiumbis(diphenylphosphino)ferrocene dichloride, dichloromethane complex (14mg, 0.0177 mmol). A carbon monoxide-filled balloon was attached and theflask was stirred in a 100 C bath for 48 hrs. The reaction was worked upby addition of water and ethyl acetate. The layers were separated andthe aqueous layer was extracted twice more with ethyl acetate. Thecombined ethyl acetate layers were washed with water and then saturatedsodium chloride solution, dried over sodium sulfate, filtered, andevaporated to a residue. The residue was purified by silica gelchromatography(methanol/dichloromethane) to give 16 mg (32%) of product.MP=98-101 C, (M+H)⁺=275.

7-(2,2-Dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazine-2-carboxylic acidcyclopentylamide. Substituting cyclopentylamine for dimethylamine.MP=237.9-241.5 C, (M+H)⁺=315.

Example 7

N-[7-(2,2-Dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-N-methyl-benzamide

Step 1: A mixture of1-[2-Bromo-5-(2-trimethylsilanyl-ethoxymethyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-one(0.25 g, 0.61 mmol), N-Methyl-benzamide (0.243 g, 1.8 mmol), K₂CO₃(0.185 g, 1.34 mmol), CuI (0.0175 g, 0.092 mmol) andN,N′-Dimethyl-ethane-1,2-diamine (0.02 mL, 0.184 mmol) in tolene washeated under an argon atmosphere in a microwave oven at 110° C.overnight. After cooling to room temperature, the reaction mixture waspurified by flash column chromatography on silica gel with EtOAc inhexane (10% to 40% gradient over 30 min) to give the desired product (60mg, 21% yield) as a yellow oil. ¹H NMR (CDCl₃, 300 MHz): δ 8.43 (s, 1H),8.04 (s, 1H), 7.45-7.24 (m, 5H), 5.65 (s, 2H), 3.74 (s, 3H), 3.62-3.59(m, 2H), 1.51 (s, 9H), 0.99-0.94 (m, 2H), 0 (s, 9H); MS [M+H]⁺: 467.Step 2: The general procedures as described in these Examples werefollowed. ¹H NMR (CDCl₃): δ 9.50 (s, 1H), 8.35 (s, 1H), 8.00 (s, 1H),7.39-7.21 (m, 5H), 3.69 (s, 3H), 1.45 (s, 9H); MS [M+H]⁺: 337.

Example 8

Cyclohexanecarboxylic acid[7-(2,2-dimethyl-propionyl)-5H-pyrrolo[2,3b]pyrazin-2-yl]-amide. Thegeneral procedures as described in these Examples were followed. ¹H NMR(DMSO-d₆): δ 12.7 (s, 1H), 10.4 (s, 1H), 9.05 (s, 1H), 8.43 (d, J=3.3Hz, 1H), 2.68-2.55 (m, 1H), 1.90-1.20 (m, 11H), 1.39 (s, 9H); MS [M+H]⁺:329.

Example 9

N-[7-(2,2-Dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-benzamide

The general procedures as described in these Examples were followed. ¹HNMR (DMSO): δ 12.85 (br s, 1H), 10.9 (s, 1H), 8.96 (S, 1H), 8.50 (s,1H), 8.14-8.12 (m, 2H), 1.62 (s) ppm, 7.765-7.53 (m, 3H), 1.40 (s, 9H);MS [M+H]⁺: 323.

Example 10

N-[7-(2,2-Dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-benzenesulfonamide

Step 1: A mixture of1-[2-Bromo-5-(2-trimethylsilanyl-ethoxymethyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-one(0.25 g, 0.61 mmol), benzenesulfonamide (0.143 g, 0.91 mmol, 1.5equiv.), CuI (0.023 g, 0.12 mmol, 20 mol %), N,N-dimethylglycine (0.013g, 0.12 mmol, 20 mol %) and K₃PO₄ (0.323 g, 1.53 mmol, 2.5 equiv.) inDMF (2 mL) was heated under an argon atmosphere in a microwave oven at153° C. overnight. After cooling to room temperature, the reactionmixture was diluted with H₂O and extracted with DCM. The combinedorganic extracts was washed with H₂O and concentrated. The residue waspurified by flash column chromatography on silica gel with EtOAc inhexane (15% to 50% gradient over 20 min) to give 0.215 g of product as apale yellow foam. ¹H NMR (CDCl₃): δ 8.50 (s, 1H), 8.29 (s, 1H),7.99-7.96 (m, 2H), 7.61-7.47 (m, 3H), 5.67 (s, 2H), 2.64-3.58 (m, 2H),1.38 (s, 9H), 1.00-0.94 (m, 2H), 0 (s, 9H); MS [M+H]⁺: 489.

Step 2: The general procedures as described in these Examples werefollowed. ¹H NMR (DMSO-d₆): δ 8.23 (s, 1H), 8.00 (s, 1H), 7.54-7.47 (m,5H), 1.17 (s, 9H); MS [M+H]⁺: 359. Example 11

N-[7-(2,2-Dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-N-methyl-benzenesulfonamide

The general procedures as described in these Examples were followedexcept replacing benzenesulfonamide with N-methylbenzenesulfonamide instep 1. ¹H NMR (CDCl₃): δ 8.75 (s, 1H), 8.49 (d, J=3.2 Hz, 1H),7.56-7.53 (m, 3H), 7.44-7.40 (m, 2H), 3.36 (s, 3H), 1.23 (s, 9H);[M+H]⁺: 373.

Example 12

2,2-dimethyl-1-(2-phenoxy-5H-pyrrolo[2,3-b]pyrazin-7-yl)-propan-1-one

A mixture of1-[2-bromo-5-(2-trimethylsilanyl-ethoxymethyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-one(0.051 g, 0.12 mmol), phenol (0.017 g, 0.18 mmol), potassium phosphate(0.056 g, 0.26 mmol),2-di-tert-butylphosphino-2′-(N,N-dimethylamino)biphenyl (0.0041 g, 0.012mmol) and Pd(OAc)₂ (0.002 g, 0.009 mmol) in 1 mL of toluene was stirredat 100° C. for 13 h, then at 150° C. for 4 h. The resulting dark orangemixture was partitioned between 10 mL of ethyl acetate and 10 mL ofwater. The organic layer was dried over MgSO₄, filtered and concentratedto an orange oil. Column chromatography (0-20% EtOAc/hexanes) afforded0.032 g (62%) of impure2,2-dimethyl-1-[2-phenoxy-5-(2-trimethylsilanyl-ethoxymethyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-propan-1-oneas a yellow oil, which was used without further purification.

Example 13

A solution of impure2,2-dimethyl-1-[2-phenoxy-5-(2-trimethylsilanyl-ethoxymethyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-propan-1-one,prepared according to general procedures as described in these Examples,(0.032 g, “0.075 mmol”) in 1 mL of dichloromethane and 1 mL oftrifluoroacetic acid was stirred for 13 h, then concentrated to ayellow-green oil, which was partitioned between 5 mL of a sat. aq.NaHCO₃ solution and 5 mL of dichloromethane. The aqueous layer wasextracted with 5 mL of dichloromethane, and the combined organic layerswere dried over MgSO₄, filtered and concentrated to a pale yellow oil.The oil was dissolved in 1 mL of ethanol and treated with sodium acetatetrihydrate (0.103 g, 0.756 mmol). The mixture was stirred for 2 h, thenconcentrated. The residue was partitioned between 5 mL of water and 5 mLof dichloromethane. The aqueous layer was extracted with 5 mL ofdichloromethane, and the combined organic layers were dried over MgSO₄,filtered and concentrated to a pale yellow oil. The oil was againdissolved in 1 mL of ethanol and treated with sodium acetate trihydrate(0.101 g, 0.745 mmol). The mixture was stirred for 4 h, thenconcentrated. The residue was partitioned between 5 mL of water and 5 mLof dichloromethane. The aqueous layer was extracted with 5 mL ofdichloromethane, and the combined organic layers were dried over MgSO₄,filtered and concentrated to an off-white solid. Column chromatography(0→33% EtOAc/hexanes) afforded 0.014 g (65% from impure; 40% 2-step) of2,2-dimethyl-1-(2-phenoxy-5H-pyrrolo[2,3-b]pyrazin-7-yl)-propan-1-one asan off-white solid.

Example 14

1-[2-(cyclopentyl-methyl-amino)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-one

A solution of1-[2-bromo-5-(2-trimethylsilanyl-ethoxymethyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-one(0.051 g, 0.123 mmol) and methylcyclopentylamine (0.050 mL, 0.44 mmol)in 1 mL of N-methyl pyrrolidinone in a sealed tube was stirred at 200°C. for 20 h. Additional methylcyclopentylamine (0.100 mL, 0.88 mmol) wasadded, and the solution stirred at 200° C. for 3 d. After cooling, thedark orange solution was partitioned between 10 mL of ethyl acetate and10 mL of a 10% citric acid solution. The organic layer was sequentiallywashed with three 10 mL portions of water and 10 mL of a sat. aq. NaClsolution, dried over MgSO₄, filtered and concentrated to an orange oil.Column chromatography (0-20% EtOAc/hexanes) afforded 0.030 g (58%) ofimpure1-[2-(cyclopentyl-methyl-amino)-5-(2-trimethylsilanyl-ethoxymethyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-one,which was used without further purification.

Example 15

A solution of impure1-[2-(cyclopentyl-methyl-amino)-5-(2-trimethylsilanyl-ethoxymethyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-one,prepared according to general procedures as described in these Examples,(0.030 g, 0.071 mmol) in 1 mL of dichloromethane and 1 mL oftrifluoroacetic acid was stirred for 3 h, then concentrated, chasingwith toluene, to a yellow oil. The oil was dissolved in 1 mL of ethanoland treated with sodium acetate trihydrate (0.103 g, 0.760 mmol). Theyellow mixture was stirred for 16 h, then concentrated to a yellowsolid. Column chromatography (two columns: first run, 0→40%EtOAc/hexanes; second run, 66% ether/hexanes) afforded 0.0045 g (21%from impure, 12% two-step) of1-[2-(cyclopentyl-methyl-amino)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-oneas a white solid.

Example 161-(2-ethoxy-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one

A mixture of7-(2,2-dimethyl-propionyl)-5-(2-trimethylsilanyl-ethoxymethyl)-1,5-dihydro-pyrrolo[2,3-b]pyrazin-2-one(0.080 g, 0.23 mmol), iodoethane (0.018 mL, 0.24 mmol) and potassiumcarbonate (0.038 g, 0.28 mmol) in 3 mL of acetone was stirred at refluxovernight, then concentrated. The residue was partitioned between 30 mLof ethyl acetate and 30 mL of water. The organic layer was dried overMgSO₄, filtered and concentred to 0.075 g (86%) of crude1-[2-ethoxy-5-(2-trimethylsilanyl-ethoxymethyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-oneas a yellow oil that was used without further purification.

Example 17

A solution of crude1-[2-ethoxy-5-(2-trimethylsilanyl-ethoxymethyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-one(0.075 g, 0.20 mmol) in 2 mL of dichloromethane and 2 mL oftrifluoroacetic acid was stirred for 3 h, then concentrated, chasingwith toluene. The resulting residue was dissolved in 3 mL of ethanol andtreated with sodium acetate trihydrate (0.270 g, 1.98 mmol). The mixturewas stirred overnight, then concentrated to a residue. Columnchromatography (0→30% EtOAc/hexanes) afforded 0.039 g (80%) of1-(2-ethoxy-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one asa white solid.

Example 18

2,2-dimethyl-1-(2-phenylamino-5H-pyrrolo[2,3-b]pyrazin-7-yl)-propan-1-one

A mixture of1-[2-bromo-5-(2-trimethylsilanyl-ethoxymethyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-one(0.206 g, 0.50 mmol), potassium carbonate (0.152 g, 1.10 mmol), copper(I) iodide (0.014 g, 0.075 mmol), proline (0.017 g, 0.15 mmol) andaniline (0.46 mL, 5 mmol) in 1 mL of dimethylsulfoxide was stirred at90° C. for 5 d, then allowed to cool. The mixture was partitionedbetween 50 mL of ethyl acetate and 30 mL of water, and the organic layerwas washed with 30 mL of a sat. aq. NaCl solution, dried over MgSO₄, andconcentrated to a residue. Column chromatography (0→45% EtOAc/hexanes)afforded 0.102 g (48%) of2,2-dimethyl-1-[2-phenylamino-5-(2-trimethylsilanyl-ethoxymethyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-propan-1-oneas a pale orange solid.

Example 19

A solution of2,2-dimethyl-1-[2-phenylamino-5-(2-trimethylsilanyl-ethoxymethyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-propan-1-one(0.102 g, 0.240 mmol) in 2 mL of dichloromethane and 1 mL oftrifluoroacetic acid was stirred for 14 h, then concentrated. Columnchromatography (20→70% EtOAc/hexanes afforded 0.055 g of theformaldehyde adduct of the desired product. This intermediate wasdissolved in 1 mL of ethanol and treated with sodium acetate trihydrate(0.327 g, 2.40 mmol). The mixture was stirred for 24 h, and solid wasisolated by filtration, rinsing with water and ethanol, and dried toafford 0.010 g (14%) of2,2-dimethyl-1-(2-phenylamino-5H-pyrrolo[2,3-b]pyrazin-7-yl)-propan-1-oneas a yellow solid.

Example 202,2-dimethyl-1-[2-(methyl-phenyl-amino)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-propan-1-one

2,2-dimethyl-1-[2-(methyl-phenyl-amino)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-propan-1-one

A mixture of1-[2-bromo-5-(2-trimethylsilanyl-ethoxymethyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-one(0.053 g, 0.129 mmol), N-methylaniline (0.021 mL, 0.19 mmol), Pd₂(dba)₃(0.012 g, 0.013 mmol), BINAP (0.012 g, 0.019 mmol) and cesium carbonate(0.084 g, 0.258 mmol) in 2 mL of toluene was stirred at 120° C. in amicrowave for 1 h, then at 150° C. in a microwave for 1 h, then at 180°C. in a microwave for 1 h, then concentrated to a residue. Columnchromatography (0→30% EtOAc/hexanes) afforded 0.021 g (37%) of2,2-dimethyl-1-[2-(methyl-phenyl-amino)-5-(2-trimethylsilanyl-ethoxymethyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-propan-1-oneas a yellow oil.

Example 21

A solution of2,2-dimethyl-1-[2-(methyl-phenyl-amino)-5-(2-trimethylsilanyl-ethoxymethyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-propan-1-one(0.021 g, 0.048 mmol) in 1 mL of dichloromethane and 1 mL oftrifluoroacetic acid was stirred for 1 h, then concentrated. The residuewas dissolved in 0.5 mL of ethanol and treated with sodium acetatetrihydrate (0.065 g, 0.48 mmol). The mixture was stirred for 15 h, thenconcentrated to a residue. Column chromatography (50→100% EtOAc/hexanes)afforded 0.009 g (61%) of2,2-dimethyl-1-[2-(methyl-phenyl-amino)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-propan-1-oneas a yellow solid.

Example 22

Cyclohexyl-3-[7-(2,2-dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-urea

A mixture of1-[2-bromo-5-(2-trimethylsilanyl-ethoxymethyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-one(0.576 g, 1.40 mmol), benzophenone imine (0.26 mL, 1.54 mmol), cesiumcarbonate (0.912 g, 2.80 mmol), palladium acetate (0.031 g, 0.14 mmol)and BINAP (0.087 g, 0.14 mmol) in 14 mL of tetrahydrofuran was stirredat 90° C. in a sealed tube for 64 h then concentrated. The resultingresidue was taken up in 30 mL of ethyl acetate and sequentially washedwith 30 mL of water and 30 mL of a sat. aq. NaCl solution, dried overMgSO₄, filtered and concentrated to a residue. Column chromatography(10→25% EtOAc/hexanes) afforded 0.670 g (93%) of1-[2-(benzhydrylidene-amino)-5-(2-trimethylsilanyl-ethoxymethyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-oneas a yellow oil.

Example 23

A mixture of1-[2-(benzhydrylidene-amino)-5-(2-trimethylsilanyl-ethoxymethyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-one(0.670 g, 1.31 mmol), sodium acetate (0.258 g, 3.14 mmol) andhydroxylamine hydrochloride (0.164 g, 2.36 mmol) in 13 mL of methanolwas stirred for 1 h, then concentrated to a residue. Columnchromatography (10→50% EtOAc/hexanes) afforded 0.359 g (79%) of1-[2-amino-5-(2-trimethylsilanyl-ethoxymethyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-oneas a white solid.

Example 24

A solution of1-[2-amino-5-(2-trimethylsilanyl-ethoxymethyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-one(0.100 g, 0.287 mmol) and cyclohexyl isocyanate (0.37 mL, 2.9 mmol) in 3mL of dichloroethane was stirred at reflux for 19 h, then concentratedto a residue. Column chromatography (10→50% EtOAc/hexanes) afforded0.103 g (76%) of1-cyclohexyl-3-[7-(2,2-dimethyl-propionyl)-5-(2-trimethylsilanyl-ethoxymethyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-ureaas a brown solid.

Example 25

A solution of1-cyclohexyl-3-[7-(2,2-dimethyl-propionyl)-5-(2-trimethylsilanyl-ethoxymethyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-urea(0.102 g, 0.215 mmol) in 2 mL of dichloromethane and 2 mL oftrifluoroacetic acid was stirred for 1 h, then concentrated, chasingtwice with toluene. The residue was dissolved in 2 mL of ethanol andtreated with sodium acetate trihydrate (0.293 g, 2.15 mmol). The mixturewas stirred for 16 h, then concentrated to a residue. Columnchromatography (50→80% EtOAc/hexanes) afforded 0.050 g (68%) of1-cyclohexyl-3-[7-(2,2-dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-ureaas a white solid.

Compound prepared in similar fashion as1-cyclohexyl-3-[7-(2,2-dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-urea:

Cycloheptyl-3-[7-(2,2-dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-urea

JAK Assay Information Determination of IC₅₀ of Janus Kinase (JAK)Inhibition:

Enzymes and peptide substrate used are described below:

-   -   JAK1: Recombinant human kinase domain from Invitrogen (Cat        #PV4774)    -   JAK3: Recombinant human kinase domain from Millipore (Cat        #14-629) or prepared.    -   JAK2: Recombinant human kinase domain from Millipore (Cat        #14-640)    -   Substrate: N-terminally biotinylated 14-mer peptide derived from        activation loop of JAK1 with sequence of the peptide substrate:        Biotin-KAIETDKEYYTVKD

Assay conditions_used are described below:

-   -   Assay Buffer: JAK Kinase Buffer: 50 mM Hepes [pH 7.2], 10 mM        MgCl₂, 1 mM DTT, 1 mg/ml BSA. The assay is carried out in this        buffer.    -   Assay Format: The kinase activity of all three JAK kinases is        measured using a radioactive, end-point assay and with trace        amounts of ³³P-ATP. The assays are carried out in 96-well        polypropylene plates.

Experimental Method:

All concentrations are final in the reaction mixture and all incubationsare carried at room temperature. Assay steps are described below:

-   -   1) Compounds are serially diluted in 100% DMSO typically at a        10× starting concentration of 1 mM. Final concentration of DMSO        in the reaction is 10%.    -   2) Compounds are preincubated with enzyme (0.5 nM JAK3        (commercially available), 0.2 nM JAK3 (prepared), 1 nM JAK2, 5        nM JAK1) for 10 minutes.    -   3) Compounds are preincubated with enzyme (0.5 nM JAK3, 1 nM        JAK2, 5 nM JAK1) for 10 minutes.    -   4) Reactions are initiated by the addition of a cocktail of the        two substrates (ATP and peptide premixed in the JAK Kinase        Buffer). In the JAK2/JAK3 assays, ATP and the peptide are used        at concentrations of 1.5 uM and 50 uM, respectively. JAK1 assay        is carried out at an ATP concentration of 10 uM and a peptide        concentration of 50 uM.    -   5) The duration of the assay for JAK2 and JAK3 is 20 minutes.        JAK1 assay is carried out for 40 minutes. With all three        enzymes, reactions are terminated by the addition of 0.5M EDTA        to a final concentration of 100 mM.    -   6) 25 ul of terminated reactions are transferred to 150 ul of a        7.5% (v/v) slurry of streptavidin-coated sepharose beads in        MgCl₂— and CaCl₂-free 1× Phosphate Buffered Saline containing 50        mM of EDTA in 96-well, 1.2 um MultiScreen-BV filter plates.    -   7) After a 30-minute incubation, the beads are washed under        vacuum with the following buffers:        -   a. 3 to 4 washes with 200 ul of 2M NaCl.        -   b. 3 to 4 washes with 200 ul of 2M NaCl plus 1% (v/v)            phosphoric acid.        -   c. 1 wash with water.    -   8) Washed plates are dried in a 60° C. oven for between 1 to 2        hours.    -   9) 70 ul of Microscint 20 scintillation fluid is added to each        well of filter plates and after at least 30 minutes of        incubation, radioactive counts are measured in a Perkinelmer        microplate scintillation counter.

Representative IC₅₀ results are in Table II below:

TABLE II IC₅₀ h-jak2- IC₅₀ h-jak3- Compound sf21-c sf21-c I-1 0.12890.0592 I-34 0.47099 I-35 0.49753

SYK Assay Information Determination of IC₅₀ of Spleen Tyrosine Kinase(SYK) Inhibition:

SYK kinase assay is a standard kinase assay adapted to a 96 well plateformat. This assay is performed in 96-well format for IC₅₀ determinationwith 8 samples which represented 10 half log dilutions and a 40 μLreaction volume. The assay measures the incorporation of radiolabeled³³P γATP into an N-terminally biotinylated peptide substrate, derivedfrom naturally occurring phosphoacceptor consensus sequence (Biotin-11aaDY*E). Phosphorylated products were detected upon termination ofreactions with EDTA and the addition of Streptavidin coated beads.Representative results are in Table II above.

-   -   Assay plates: 96-well MultiScreen 0.65 um filter plates        (Millipore Cat. No.: MADVNOB10)    -   Streptavidin coated beads: Streptavidin Sepharose™, suspension        5.0 mL, in 50 mM EDTA/PBS diluted (1:100), (Amersham, Cat. No.:        17-5113-01)    -   Compounds: 10 mM in 100% dimethylsulfoxide (DMSO), final conc.:        compound 0.003-100 uM in 10% DMSO    -   Enzyme: SYK RPA purified, truncated construct of Spleen Tyrosine        Kinase aa 360-635, stock solution 1 mg/mL, MW: 31.2 KDa, final        conc.: 0.0005 μM.    -   Peptide 1: biotinylated peptide is derived from a naturally        occurring phosphor-acceptor consensus sequence        (Biotin-EPEGDYEEVLE), special order from QCB, stock solution 20        mM, final conc.: 5.0 μM.    -   ATP: Adenosine-5′-triphosphate 20 mM, (ROCHE Cat. No.:        93202720), final concentration: 20 μM    -   Buffer: HEPES: 2-Hydroxyethyl piperazine-2-ethanesulfonic acid        (Sigma, Cat. No.: H-3375) final concentration: 50 mM HEPES pH7.5    -   BSA: Bovine Serum Albumin Fraction V, fatty acid free (Roche        Diagnostics GmbH, Cat. No. 9100221) diluted to a final        concentration of 0.1%    -   EDTA: EDTA stock solution 500 mM, (GIBCO, Cat. No.: 15575-038)        final concentration: 0.1 mM    -   DTT: 1,4-Dithiothreitol (Roche Diagnostics GmbH, Cat. No.:        197777), final conc.: 1 mM    -   MgCl₂×6H₂O: MERCK, Cat. No.: 105833.1000, final concentration:        10 mM Assay Dilution Buffer (ADB): 50 mM HEPES, 0.1 mM EGTA, 0.1        mM Na Vanadate, 0.1 mM β-glycerophosphate, 10 mM MgCl₂, 1 mM        DTT, 0.1% BSA, pH 7.5    -   Bead wash buffer: 10 g/L PBS (Phosphate buffered saline) with 2M        NaCl+1% phosphoric acid.

Experimental Method:

In 40 μL volume, 26 μL of ADB diluted, purified recombinant humanSYK360-635 [0.5 nM] was mixed with 4 μL of 10× concentrations of thetest compounds, [usually 100 μM-0.003 μM] in [10%] DMSO and the mixturewas incubated for 10 min at RT.

The kinase reaction was initiated by the addition of 10 μL 4× substratecocktail containing the DYE peptide substrate [0 or 5 μM], ATP [20 μM]and ³³PγATP [2 μCi/rxn]. After incubation at 30° C. for 15 min, thereaction was terminated by the transfer of 25 μL pf the reaction sampleto a 96 well 0.65 μm Millipore MADVNOB membrane/plate containing 200 μL5 mM EDTA and 20% Streptavidine coated beads in PBS.

The unbound radionucleotides were washed under vacuum with 3×250 μL 2MNaCl; 2×250 μL 2M NaCl+1% phosphoric acid; 1×250 μL H₂O. After the lastwash membrane/plates were transferred to an adaptor plate, heat driedfor 15 min at 60° C., and 50 μL scintillation cocktail was added to eachwell and 4 h later the amount of radioactivity was counted in a topcounter.

The percent inhibition was calculated based on the uninhibited enzymerate:

% Inhibition=100/(1+(IC ₅₀/Inhibitor conc)^(n))

-   -   The IC₅₀ was calculated using a non-linear curve fit with XLfit        software (ID Business Solution Ltd., Guilford, Surrey, UK).

The foregoing invention has been described in some detail by way ofillustration and example, for purposes of clarity and understanding. Itwill be obvious to one of skill in the art that changes andmodifications may be practiced within the scope of the appended claims.Therefore, it is to be understood that the above description is intendedto be illustrative and not restrictive. The scope of the inventionshould, therefore, be determined not with reference to the abovedescription, but should instead be determined with reference to thefollowing appended claims, along with the full scope of equivalents towhich such claims are entitled.

All patents, patent applications and publications cited in thisapplication are hereby incorporated by reference in their entirety forall purposes to the same extent as if each individual patent, patentapplication or publication were so individually denoted.

1. A compound of Formula I

R is R¹, R², R³, or R⁴; R¹ is lower alkyl, lower alkoxy, phenyl, benzyl,heteroaryl, cycloalkyl, heterocycloalkyl, or cycloalkylalkyl, optionallysubstituted with one or more R^(1a); R^(1a) is R^(1b) or R^(1c); R^(1b)is halogen, oxo, hydroxy, or —CN; R^(1c) is —C(═O)O(R^(1f)),—C(═O)CH₂(R^(1e)), —S(R^(1f)), —S(O)₂(R^(1f)), or —S(═O)(R^(1f)), loweralkyl, lower alkoxy, amino, amido, lower haloalkyl, phenyl, heteroaryl,cycloalkyl, heterocycloalkyl, cycloalkyloxy, or heterocycloalkyloxyoptionally substituted with one or more R^(1d); R^(1d) is H, halogen,hydroxy, lower alkyl, lower alkoxy, or lower haloalkyl; R^(1e) is H,lower alkyl, lower alkoxy, —CN, lower haloalkyl, phenyl, heteroaryl,cycloalkyl, or heterocycloalkyl; R^(1f) is H, lower alkyl, lowerhaloalkyl, phenyl, heteroaryl, cycloalkyl, or heterocycloalkyl; R² isN(R^(2a))₂; each R^(2a) is independently H or R^(2b); each R^(2b) isindependently lower alkyl, phenyl, heteroaryl, cycloalkyl,heterocycloalkyl, or heterocycloalkyl alkylene, optionally substitutedwith one or more R^(2c); R^(2c) is R^(2d) or R^(2e);  R^(2d) is halogen,oxo, or hydroxy;  R^(2e) is —N(R^(2g))₂, —C(═O)(R^(2g)),—C(=O)O(R^(2g)), —C(═O)N(R^(2g))₂, —N(R^(2g))C(═O)(R^(2g)),—S(═O)₂(R^(2g)), —S(O)₂N(R^(2g))₂, lower alkyl, lower alkoxy, lowerhaloalkyl, phenyl, heteroaryl, heteroaryloxy, cycloalkyl, orheterocycloalkyl, optionally substituted with one or more R^(2f);  eachR^(2f) is independently H, halogen, lower alkyl, lower alkoxy, lowerhaloalkyl;  each R^(2g) is independently H, lower alkyl, lower alkoxy,lower haloalkyl, or phenyl; R³ is —C(═O)R^(3a); R^(3a) is lower alkyl,lower alkoxy, phenyl, or N(R^(3b))₂; each R^(3b) is independently H orlower alkyl; R⁴ is —O(R^(4a)); R^(4a) is H or R^(4b); R^(4b) is loweralkyl, phenyl, benzyl, lower haloalkyl, cycloalkyl, heterocycloalkyl,heteroaryl, optionally substituted with one or more R^(4c); R^(4c) ishalogen, hydroxy, lower alkyl, lower haloalkyl, or lower alkoxy; Q³ is—O-Q^(3a), —S-Q^(3a), —C(═O)(Q^(3a)), —O(CH₂)_(m)C(═O)(Q^(3a)),—S(═O)(Q^(3a)), —S(═O)₂(Q^(3a)), —N(Q^(3a))₂, —N(Q^(3a))S(═O)₂(Q^(3a)),—N(Q^(3a))C(═O)(Q^(3a)), —C(═O)N(Q^(3a))₂, or —N(Q^(3a))C(═O)N(Q^(3a))₂;each Q^(3a) is independently Q^(3b) or Q^(3c); m is 0, 1, or 2; Q^(3b)is H; Q^(3c) is lower alkyl, lower haloalkyl, phenyl, cycloalkyl,heterocycloalkyl, or heteroaryl, optionally substituted with one or moreQ^(3d); and each Q^(3d) is independently Q^(3d) is Q^(3e) or Q^(3f);Q^(3e) is halogen or hydroxy; Q^(3f) is lower alkyl, lower alkoxy, lowerhaloalkyl, phenyl, cycloalkyl, heterocycloalkyl, or heteroaryl,optionally substituted with one or more Q^(3g); and each Q^(3g) isindependently halogen, hydroxy, lower alkyl, lower hydroxyalkyl, lowerhaloalkyl, or lower alkoxy; or a pharmaceutically acceptable saltthereof.
 2. The compound of claim 1, wherein R is R¹.
 3. The compound ofclaim 2, wherein R¹ is lower alkyl.
 4. The compound of claim 3, whereinR¹ is tert-butyl.
 5. The compound of claim 3, wherein R¹ is —CHC(CH₃)₃.6. The compound of claim 3, wherein R¹ is iso-butyl.
 7. The compound ofclaim 3, wherein R¹ is iso-propyl.
 8. The compound of claim 2, whereinR¹ is cycloalkyl.
 9. The compound of claim 2, wherein R¹ isheterocycloalkyl.
 10. The compound of claim 2, wherein R¹ is benzyl. 11.The compound of claim 2, wherein R¹ is phenyl.
 12. The compound of claim1, wherein R is R².
 13. The compound of claim 12, wherein R² isNH(R^(2a)).
 14. The compound of claim 13, wherein R^(2a) is R^(2b) andR^(2b) is lower alkyl.
 15. The compound of claim 14, wherein R^(2b) isiso-propyl.
 16. The compound of claim 13, wherein R^(2a) is R^(2b) andR^(2b) is heterocycloalkyl.
 17. The compound of claim 13, wherein R^(2a)is R^(2b) and R^(2b) is cycloalkyl.
 18. The compound of claim 13,wherein R^(2a) is R^(2b) and R^(2b) is heterocycloalkyl alkylene. 19.The compound of claim 18, wherein R^(2b) is pyrrolidinyl alkylene. 20.The compound of claim 18, wherein R^(2b) is pyrrolidinyl methylene. 21.A compound selected from the group consisting of:2-(Cyclopentyl-methyl-amino)-5H-pyrrolo[2,3-b]pyrazine-7-carboxylic acidisopropylamide;1-[7-(2,2-Dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-2,2-dimethyl-propan-1-one;1-(2-Cyclopentyloxy-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one;N-[7-(2,2-Dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-benzamide;Cyclohexanecarboxylic acid[7-(2,2-dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-amide;N-[7-(2,2-Dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-N-methyl-benzamide;N-[7-(2,2-Dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-benzenesulfonamide;N-[7-(2,2-Dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-N-methyl-benzenesulfonamide;7-(2,2-Dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazine-2-carboxylic aciddimethylamide;7-(2,2-Dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazine-2-carboxylic acidisopropylamide;1-(2-Acetyl-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one;1-(2-Isobutyryl-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one;2,2-Dimethyl-1-[2-(2-methyl-benzoyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-propan-1-one;1-(2-Benzoyl-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one;7-(2,2-Dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazine-2-carboxylic acidcyclopentylamide;2,2-Dimethyl-1-[2-(pyridine-4-carbonyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-propan-1-one;2,2-Dimethyl-1-[2-(pyridine-3-carbonyl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-propan-1-one;1-(2-Cycloheptyloxy-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one;1-(2-Cyclohexyloxy-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one;1-(2-Isopropoxy-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one;1-(2-Ethoxy-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one;2,2-Dimethyl-1-[2-(2-morpholin-4-yl-2-oxo-ethoxy)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-propan-1-one;1-[2-(1,2-Dimethyl-propoxy)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-one;1-(2-Isobutoxy-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one;1-[2-(1-Benzyl-1H-[1,2,3]triazol-4-yl)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-one;1-Cyclohexyl-3-[7-(2,2-dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-urea;1-Cycloheptyl-3-[7-(2,2-dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-urea;2,2-Dimethyl-1-[2-(methyl-phenyl-amino)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-propan-1-one;1-(2-sec-Butoxy-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one;2,2-Dimethyl-1-(2-phenylamino-5H-pyrrolo[2,3-b]pyrazin-7-yl)-propan-1-one;1-(2-Cyclopentyloxy-5H-pyrrolo[2,3-b]pyrazin-7-yl)-2,2-dimethyl-propan-1-one;1-[2-(Cyclopentyl-methyl-amino)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-one;2,2-Dimethyl-1-(2-phenoxy-5H-pyrrolo[2,3-b]pyrazin-7-yl)-propan-1-one;2,2-Dimethyl-1-{2-[4-(4-methyl-piperazin-1-yl)-phenylamino]-5H-pyrrolo[2,3-b]pyrazin-7-yl}-propan-1-one;1-Cyclohexyl-3-[7-(2,2-dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-urea;1-Cycloheptyl-3-[7-(2,2-dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-urea;1-[7-(2,2-Dimethyl-propionyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl]-2,2-dimethyl-propan-1-one;and1-[2-(6-Methoxy-pyridin-2-ylamino)-5H-pyrrolo[2,3-b]pyrazin-7-yl]-2,2-dimethyl-propan-1-one.22. A method for treating an inflammatory or autoimmune conditioncomprising administering to a patient in need thereof a therapeuticallyeffective amount of the compound of claim
 1. 23. The method of claim 22,further comprising administering an additional therapeutic agentselected from a chemotherapeutic or anti-proliferative agent, ananti-inflammatory agent, an immunomodulatory or immunosuppressive agent,a neurotrophic factor, an agent for treating cardiovascular disease, anagent for treating diabetes, or an agent for treating immunodeficiencydisorders.
 24. A method for treating an inflammatory conditioncomprising administering to a patient in need thereof a therapeuticallyeffective amount of the compound of claim
 1. 25. A method for inhibitingT-cell proliferative disorder comprising administering to a patient inneed thereof a therapeutically effective amount of the compound ofclaim
 1. 26. The method of claim 25, wherein the proliferative disorderis cancer.
 27. A method for treating a B-cell proliferative disordercomprising administering to a patient in need thereof a therapeuticallyeffective amount of the compound of claim
 1. 28. A method for treatingan immune disorder including lupus, multiple sclerosis, rheumatoidarthritis, psoriasis, Type I diabetes, complications from organtransplants, xeno transplantation, diabetes, cancer, asthma, atopicdermatitis, autoimmune thyroid disorders, ulcerative colitis, Crohn'sdisease, Alzheimer's disease, and Leukemia, comprising administering toa patient in need thereof a therapeutically effective amount of thecompound of claim
 1. 29. A method for preventing or treating all formsof organ rejection, including acute allograft or xenograft rejection andchronic allograft or xenograft rejection, of vascularized ornon-vascularized transplants, comprising administering to a patient inneed thereof the compound of claim
 1. 30. A method for inhibiting JAK3activity comprising administering the compound of claim 1, wherein thecompound exhibits an IC₅₀ of 50 micromolar or less in an in vitrobiochemical assay of JAK3 activity.
 31. The method of claim 30 whereinthe compound exhibits an IC₅₀ of 100 nanomolar or less in an in vitrobiochemical assay of JAK3 activity.
 32. The method of claim 31 whereinthe compound exhibits an IC₅₀ of 10 nanomolar or less in an in vitrobiochemical assay of JAK3 activity.
 33. A method for inhibiting SYKactivity comprising administering the compound of claim 1, wherein thecompound exhibits an IC₅₀ of 50 micromolar or less in an in vitrobiochemical assay of SYK activity.
 34. The method of claim 33 whereinthe compound exhibits an IC₅₀ of 100 nanomolar or less in an in vitrobiochemical assay of SYK activity.
 35. The method of claim 34 whereinthe compound exhibits an IC₅₀ of 10 nanomolar or less in an in vitrobiochemical assay of SYK activity.
 36. A method for treating aninflammatory condition comprising co-administering to a patient in needthereof a therapeutically effective amount of an anti-inflammatorycompound in combination with the compound of claim
 1. 37. A method fortreating an immune disorder comprising co-administering to a patient inneed thereof a therapeutically effective amount of an immunosuppressantcompound in combination with the compound of claim
 1. 38. Apharmaceutical composition comprising the compound of claim 1, admixedwith at least one pharmaceutically acceptable carrier, excipient ordiluent.
 39. The pharmaceutical composition of claim 38, furthercomprising an additional therapeutic agent selected from achemotherapeutic or anti-proliferative agent, an anti-inflammatoryagent, an immunomodulatory or immunosuppressive agent, a neurotrophicfactor, an agent for treating cardiovascular disease, an agent fortreating diabetes, and an agent for treating immunodeficiency disorders.40. Use of the compound of claim 1 in the manufacture of a medicamentfor the treatment of an inflammatory disorder.
 41. Use of the compoundof claim 1 in the manufacture of a medicament for the treatment of anautoimmune disorder.